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The Connective Tissue Growth Factor/Cysteine- Rich 61/Nephroblastoma Overexpressed (CCN) Family¹

Departments of Surgery and Medical Biochemistry, The Ohio State University, Columbus, Ohio 43210; and Department of Surgery, Children’s Hospital, Columbus, Ohio 43205

	Abstract

Top Abstract I. Introduction II. The Modular Structure... III. Connective Tissue Growth... IV. Cyr61 V. Nov VI. Elm1/WISP-1 VII. Heparin-Induced CCN-Like... VIII. WISP-3 IX. Other CCN-Like Molecules X. Regulation of Cellular... XI. Biological Processes... XII. Perspectives and Future... References

 

I. Introduction

II. The Modular Structure of the Connective Tissue Growth Factor/Cysteine-Rich 61/Nephroblastoma Overexpressed (CCN) Family Members

III. Connective Tissue Growth Factor (CTGF)

A. Discovery

B. Structure of the CTGF gene and protein

C. CTGF mRNA production

D. CTGF production, secretion, and processing

E. CTGF-heparin interactions

F. Biological properties of CTGF

G. Mechanism of action of CTGF

IV. Cyr61

A. Discovery of cyr61

B. Structure of the cyr61 gene and protein

C. Cyr61 mRNA production

D. Cyr61 production and secretion

E. Biological properties of cyr61

F. Mechanism of action of cyr61

V. Nov

A. Discovery and structure of nov

B. Nov mRNA production

C. Biological properties of nov

VI. Elm1/WISP-1

VII. Heparin-Induced CCN-Like Protein (HICP)/rCop-1/CTGF-3/WISP-2

VIII. WISP-3

IX. Other CCN-Like Molecules

X. Regulation of Cellular Functions by the CCN Family

A. Cell cycle control

B. Cell adhesion and migration

C. Extracellular matrix (ECM) production

XI. Biological Processes Involving the CCN Family

A. Development and differentiation

B. Female reproductive tract function

C. Angiogenesis

D. Wound repair

E. Fibrotic disorders

F. Inflammation

G. Tumor growth

XII. Perspectives and Future Directions

	I. Introduction

Top Abstract I. Introduction II. The Modular Structure... III. Connective Tissue Growth... IV. Cyr61 V. Nov VI. Elm1/WISP-1 VII. Heparin-Induced CCN-Like... VIII. WISP-3 IX. Other CCN-Like Molecules X. Regulation of Cellular... XI. Biological Processes... XII. Perspectives and Future... References

 
THE LAST 5–6 yr have seen the emergence of a new gene family that currently comprises connective tissue growth factor (CTGF; also termed fisp-12), cysteine-rich 61 (cyr61), nephroblastoma overexpressed (nov), expressed low in metastasis 1 (elm1; also termed WISP-1), heparin-inducible CTGF/cyr61/nov (CCN)-like protein (HICP; also termed rCop-1, CTGF-3 or WISP-2), and WISP-3 (Fig. 1). Family members have been characterized from human, mouse, rat, pig, cow, chicken, quail, and frog and are predicted to have arisen from a common ancestral gene more than 40 million years ago (Fig. 2). Although these proteins were initially classified as immediate early gene products or growth factors, this concept has had to be modified in light of more detailed studies of their activities as well as the discovery of unique family members that exhibit quite different biological properties. Since CTGF, cyr61, and nov were the prototype members of this family, this article adopts the term "CCN family" as introduced by Bork in 1993 (1). Implicit in this usage is that reference to the CCN family applies to all paralogs, and that CTGF, cyr61, and nov are not necessarily representative of the full range of gene or protein structures and biological properties.

View larger version (96K): [in this window] [in a new window]   Figure 1. Amino acid sequence alignment of human CCN paralogs. Sequences were aligned using the J. Hein method. Shading represents residues that are identical to the consensus sequence.

View larger version (11K): [in this window] [in a new window]   Figure 2. Dendrogram of human CCN paralogs. The x-axis is millions of years.

	II. The Modular Structure of Connective Tissue Growth Factor/Cysteine Rich 61/Nephroblastoma Overexpressed (CCN) Family Members

Top Abstract I. Introduction II. The Modular Structure... III. Connective Tissue Growth... IV. Cyr61 V. Nov VI. Elm1/WISP-1 VII. Heparin-Induced CCN-Like... VIII. WISP-3 IX. Other CCN-Like Molecules X. Regulation of Cellular... XI. Biological Processes... XII. Perspectives and Future... References

 
Through evolution, exons with specific biological functions ("modules") have been shuffled, forming genes that encode mosaic proteins which exhibit new biological properties (2, 3, 4). CTGF, cyr61, and nov were shown to contain four distinct structural modules that exhibit homology to conserved regions in a variety of extracellular mosaic proteins (1). All four modules are represented in WISP-1 and WISP-3, and three of the modules are present in rCop-1 and its orthologs (Fig. 3). Each module is involved in protein binding and contains conserved cysteine, hydrophobic, and polar residues. Except for module 2 in WISP-3, the modules in the various CCN proteins are 38–98% conserved with the corresponding module in human CTGF (hCTGF) (Table 1). While complex intrachain disulfide bridging is likely, this is predicted to occur within, rather than between, the modules (1).

View larger version (33K): [in this window] [in a new window]   Figure 3. Modular structure of individual CCN family members. Proteins shown with black boxes are the predicted primary translational products. Proteins shown with open boxes are biologically active derivatives arising by proteolysis (CTGF) or viral DNA integration (Nov). Question marks indicate that the C termini of N-terminally truncated CTGF proteins have not been experimentally determined.

Module 1 is approximately 32% identical with the N-terminal cysteine-rich regions of the six "classic" IGF-binding proteins, IGFBP-1 to -6 (1), and contains a motif (GCGCCXXC) that is involved in binding IGF (17). ¹²⁵I-labeled IGF-I or -II specifically binds recombinant hCTGF (rhCTGF) (12), although with much lower affinity than classic IGFBPs and more comparable to mac25, a low-affinity IGFBP, also termed IGFBP-7 or IGFBP-related protein (IGFBP-rP)-1 (18). Nov was also reported to bind IGFs (19), but this result has not proven reproducible (20). Recently, the terms IGFBP-8, -9, and -10 were introduced as synonyms for CTGF, nov, and cyr61, respectively (12), although it has since been proposed to use the names IGFBP-rP-2, -3, and -4 until their relationship to the classic IGFBPs has been determined (21). Indeed, a case against classifying the CCN family as IGFBPs has recently been made (22). Whatever the terminology, it remains unclear whether the interaction of IGFs with CCN family members occurs physiologically, whether sequences within module 1 are actually responsible for the observed binding of IGF, and what consequences this interaction has on the respective half-life, bioavailability, or activity of each of the molecules involved. However, there is general agreement that direct, IGF-independent effects of CTGF on the cell cycle are likely to be more significant than its low affinity binding of IGF (12, 22).

Module 2 comprises a Von Willebrand type C domain (VWC) that occurs in Von Willebrand factor as well as various mucins, thrombospondins, and collagens (1). Many proteins that contain VWC modules participate in oligomerization, which may be preceded by a dimerization event (23). Since module 4 of CTGF is a putative dimerization domain, module 2 may mediate the formation of complexes from CTGF dimers. Unlike all other CCN proteins, module 2 in WISP-3 contains only 6 of the 10 cysteines, the functional significance of which has yet to be established (24). Module 3 is a thrombospondin type 1 (TSP1) that contains the local motif WSXCSXXCG (1) and appears to be a cell attachment motif that binds sulfated glycoconjugates (1, 25, 26, 27). While cyr61 and CTGF promote cell adhesion (15, 16), the role of module 3 in this process remains unexplored.

Module 4 is a C-terminal (CT) module that also occurs in the C termini of a variety of unrelated extracellular mosaic proteins (1). Six of the 10 cysteine residues in the CT module appear to adopt the cystine knot motif that also occurs in nerve growth factor (NGF), transforming growth factor-ß (TGF-ß), and platelet-derived growth factor (PDGF) (1). This complex structure comprises two 2-stranded ß-sheets that lie face-to-face and are linked by three interlocking disulfide bridges and has defined a new superfamily of growth regulators (28, 29) to which members of the CCN family (except orthologs of rCop-1, which lack module 4; see below) may also belong. Since some of the receptor binding properties of NGF, TGF-ß, and PDGF reside in variable regions within the cystine knot (28), the CT module likely contains both dimerization and receptor-binding domains (1). Although dimerization of CCN proteins has not been reported, bioactive forms of 10-kDa porcine CTGF (pCTGF) comprise the C-terminal 102 or 103 residues of the primary translational product (Fig. 3) (10) and thus support the proposed role of module 4 in binding cell surface receptors (1). Module 4 is absent from rCop and its orthologs (Figs. 1 and 3), suggesting that they are functionally distinct from other CCN proteins.

	III. Connective Tissue Growth Factor (CTGF)

Top Abstract I. Introduction II. The Modular Structure... III. Connective Tissue Growth... IV. Cyr61 V. Nov VI. Elm1/WISP-1 VII. Heparin-Induced CCN-Like... VIII. WISP-3 IX. Other CCN-Like Molecules X. Regulation of Cellular... XI. Biological Processes... XII. Perspectives and Future... References

 
A. Discovery
The first description of a CTGF ortholog occurred in 1988 when cDNA encoding "fibroblast-inducible secreted protein-12" (fisp-12) was isolated by differential screening of a cDNA library from serum-stimulated NIH 3T3 cells (30). The gene structure and predicted protein sequence of fisp-12 were subsequently reported in 1991 (5). Using a similar screening strategy, Brunner et al. (31) independently isolated the same cDNA, termed ßIG-M2, from TGF-ß2-stimulated mouse AKR-2B cells. Fisp-12/ßIG-M2 comprises 348 amino acids and contains 39 Cys residues, one of which occurs in the 25-residue N-terminal signal peptide (Fig. 1) (5, 31). The human ortholog of CTGF was discovered in 1991 due to the cross-reactivity of a PDGF antiserum with 38-kDa hCTGF secreted by cultured human vein endothelial cells (HUVECs) (13). The corresponding cDNA was isolated by screening a HUVEC cDNA expression library with anti-PDGF and shown to encode a 349-amino acid protein that is 91% homologous to fisp-12 (13) (Fig. 1). The protein was termed "CTGF" because it was both mitogenic and chemotactic for fibroblast-like cells in vitro. CTGF cDNAs have since been reported for pig, rat, cow, and frog (10, 32, 33, 34, 35).

B. Structure of the CTGF gene and protein
The fisp-12 gene comprises five exons and four introns and spans 3.1 kb (Ref. 5 ; Table 2). The organization and structure of the hCTGF gene are very similar to that of fisp-12 except that exon 1 encodes one additional amino acid in the signal peptide (13, 36, 37). There is 80% sequence identity between CTGF and fisp-12 in the 300 nucleotides that lie immediately upstream of the mRNA cap site (36), and the 5'-regions of both genes contain a variety of conventional regulatory elements as well as a unique TGF-ß response element (Fig. 4). Fisp-12 maps to the A3-B1 region of murine chromosome 10 (5) and hCTGF maps to human chromosome 6q23.1 (38).

View larger version (14K): [in this window] [in a new window]   Figure 4. Key regulatory elements in the gene promoters of hCTGF, mouse cyr61, and human nov. Note that the TGF-ß response element (TßRE) is unique to CTGF.

C. CTGF mRNA production
Among the cell types that produce CTGF mRNA are fibroblasts (5, 14, 31, 35, 39, 41, 42), endothelial cells (13, 35, 43), vascular smooth muscle cells (VSMC) (35, 44), epithelial cells (45, 46, 47), chondroctyes (48), and glioblastoma cells (37). Fisp-12/CTGF transcripts have been detected in multiple tissues of at least four mammalian species (5, 12, 32, 35, 47, 49). Most studies have reported a single CTGF transcript of 2.4 kb, although glioblastoma cells also contain 3.5-kb and 7.0-kb transcripts (37). The 2.4-kb transcript is present in unstimulated fibroblasts (35, 39) but is rapidly induced after treatment with TGF-ß or serum and is superinduced in stimulated fibroblasts in the absence of de novo protein synthesis (5, 31, 41). These studies collectively established that CTGF/fisp-12 are encoded by immediate early genes, although their kinetics of induction are more rapid and sustained than those of other immediate early genes (41). TGF-ß also increases CTGF mRNA levels in cultured human or mouse lung mesenchymal cells (34) and in human chondrocytic cells (48), the latter of which also produce CTGF mRNA in response to bone morphogenic protein-2, a member of the TGF-ß family (50). Using an enzyme-linked immunosorbent assay (ELISA), TGF-ß treatment of human fibroblasts was shown to result in increased production of the CTGF protein (51). The molecular basis for the action of TGF-ß on CTGF gene transcription has been atributed to nucleotides -157 to -145 of the hCTGF promoter (36). This region is a unique TGF-ß-inducible element (Fig. 4) that is fully conserved in the promoter of the fisp-12 gene (5) but is absent from the other parologs of the gene family (Fig. 4) as well as from other immediate early genes. In addition, inhibitors of protein kinase A, but not of tyrosine kinases or protein kinase C, block TGF-ß-stimulated CTGF transcription (52). Collectively, the TGF-ß response element and the involvement of cAMP in CTGF mRNA production represent major mechanistic differences in CTGF gene transcription as compared with other CCN family members.

Although considerable attention has been directed toward the TGF-ß inducibility of CTGF expression and the potential pathophysiological consequences thereof (53), PDGF, epidermal growth factor (EGF), and fibroblast growth factor (FGF) also stimulated hCTGF gene expression in fibroblasts (41). As compared with the effects of TGF-ß on this cell type, CTGF transcription is less robust in response to these growth factors (41) and does not result in increased CTGF protein levels (51). In contrast, pancreatic cancer cells of epithelial origin demonstrate much stronger production of CTGF mRNA by EGF or TGF- than TGF-ß, although the lack of response to TGF-ß was due, at least partly, to defects in TGF-ß signaling (54). Phenomenologically, the lack of response to TGF-ß in certain pancreatic cells is similar to that of skin epithelial cells, which (unlike fibroblasts) do not synthesize CTGF in response to local TGF-ß administration in vivo (14). Dexamethasone treatment of cultured 3T3 cells was recently shown to stimulate CTGF expression while actually down-regulating TGF-ß mRNA (55). Systemic administration of dexamethasone stimulated CTGF expression in the heart, kidney, and skin. Basal and dexamethasone-stimulated CTGF expression was strongly attenuated by tumor necrosis factor- (TNF-) (55), which was independently shown to suppress basal CTGF mRNA production in bovine aortic endothelial cells (BAECs), fibroblasts, and VSMCs (35). Finally, increased CTGF expression in renal epithelial cells in response to injury occurred without an increase in TGF-ß expression (46). These results show, not surprisingly, that mechanisms of CTGF gene regulation may be cell specific and, in addition to the action of TGF-ß, involve a variety of hormones, growth factors, and cytokines.

D. CTGF production, secretion, and processing
CTGF is present in HUVEC-conditioned medium (13) and undergoes microsomal processing (5), consistent with the presence of a signal peptide and its presumed secretion. CTGF mRNA is expressed in BAECs in which it is present at higher levels during growth than confluence (43). Similarly, expression of fisp-12 in cycling fibroblasts is relatively high (15). CTGF produced by human foreskin fibroblasts or mouse connective tissue cells under normal growth conditions appears to be cell associated, long lived, and secreted relatively inefficiently (39). On the other hand, fisp-12 was secreted efficiently from NIH 3T3 cells but was relatively unstable in the medium (5). Kireeva et al. (15) showed that fisp-12 was present in the cellular fraction, ECM, and medium and that inhibition of lysosomal acid hydrolysis increased the half-life of fisp-12 in the cellular fraction and stabilized steady-state fisp-12 levels. These data suggest that membrane-associated fisp-12 may form complexes with receptors on the surface of fisp-12-producing cells that are then internalized and degraded (15).

Various low mass (10–20 kDa) forms of pCTGF were purified from uterine secretory fluids and shown to commence between modules 2 and 3, at the beginning of module 3, or between modules 3 and 4 (10, 11) (Fig. 3). Native 38-kDa pCTGF undergoes rapid proteolytic processing by uterine fluids (11) and likely explains the presence of 38-kDa CTGF in uterine tissues (32) but not in uterine fluids (10, 11). Native low-mass CTGFs in uterine fluids are extremely stable, and their levels are strongly correlated with those of CTGF-degrading proteases (11). Although conditioned medium from serum-stimulated mouse fibroblasts did not degrade fisp-12 (15), 10- to 12-kDa CTGF, but not 38-kDa CTGF, was present in conditioned medium from cycling human or mouse fibroblasts (39). Recently, Western blot analysis demonstrated 12- to 14-kDa C-terminal forms of CTGF in mouse uterine luminal flushings (47) and 18- and 24-kDa CTGF isoforms in human serum and amniotic, follicular, peritoneal, and cerebrospinal fluids (40). The presence of CTGF in sera has been confirmed by ELISA (51). Whereas serum CTGF levels in normal human subjects were less than 28 ng/ml, those of patients with biliary atresia were up to 8-fold higher (51). These results highlight potential diagnostic or prognostic applications of measuring circulating or secreted CTGF levels.

Collectively, these data suggest that qualitative and quantitative aspects of CTGF secretion may be regulated by numerous factors including stage of cell cycle, growth factor pretreatment, cell type, species and tissue of origin, and protease activity of the pericellular environment. Moreover, the presence of CTGF in a variety of body fluids suggests that they may be important reservoirs of CTGF in vivo.

E. CTGF-heparin interactions
Heparin-affinity chromatography has been used to purify 38-kDa rhCTGF (14) and 10- to 20-kDa pCTGF (10, 11), the latter of which are eluted from heparin by 0.8 M NaCl. ³⁵S-labeled fisp-12 binds to heparin-agarose beads and is eluted by 0.4 M NaCl (15). Although module 3 is a binding motif for sulfated glycoconjugates (1), residues 247–260, 274–286, and 305–328 of hCTGF bind strongly to heparin (10, 56). Residues 247–260 contain a proposed heparin-binding consensus sequence (XBBXBX) that occurs in a variety of heparin-binding proteins (57).

As with other heparin-binding growth factors, such as FGFs, heparin-binding EGF-like growth factor, amphiregulin, keratinocyte growth factor, and vascular endothelial growth factor (58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70), the biological activity of CTGF is modulated by heparin (10, 14). The binding of fisp-12 to heparin has been proposed as the basis for the association of fisp-12 with the ECM in cultured cells (15). Heparin-like molecules on the cell surface or in the ECM may constitute a high-capacity, low-affinity binding reservoir for CTGF and thereby regulate its activity, bioavailability, or stability. Indeed, the half-life of fisp-12 in ECM (1 h) is much less than that of cyr61 (4 h) and appears to correlate with the relative affinity of each paralog for heparin (15, 71).

F. Biological properties of CTGF
Native 38-kDa hCTGF is a mitogenic and chemotactic factor for NIH 3T3 cells in vitro (13). Ten-kilodalton forms of pCTGF are mitogenic for Balb/c 3T3 cells, VSMCs, and endometrial stromal cells, but not endothelial cells (10, 56). The mitogenic activity of 38-kDa rhCTGF or 10-kDa pCTGF on fibroblasts is enhanced by EGF, PDGF, basic FGF (bFGF), or IGF-I and is either enhanced or reduced by heparin according to the CTGF-heparin ratio (10, 14). In monolayer cultures of normal rat kidney (NRK) cells, 38-kDa rhCTGF stimulated DNA synthesis and induced expression of type 1 collagen, fibronectin, and 5 integrin (14), as is characteristic of the effects of TGF-ß. These various biological effects are elicted by CTGF concentrations of about 1–20 ng/ml. In vivo, injection of TGF-ß or CTGF into the dermal/subcuticular area of the skin in neonatal mice produced nodules comprising mainly connective tissue cells and ECM (14, 72), suggesting that CTGF plays a role in TGF-ß-mediated formation of granulation tissue. Recombinant hCTGF is also mitogenic for human lung fibroblasts (73). However, recombinant 38 kDa fisp-12 did not directly stimulate DNA synthesis in HUVECs or 3T3 cells at 0.3–3 µg/ml (15), although other evidence suggests that endothelial cells do not respond mitogenically to 10-kDa CTGF, which is nonetheless mitogenic for 3T3 cells and VSMCs (10). At 0.3–3 µg/ml, the same fisp-12 protein enhanced the mitogenic activity of 10 ng/ml bFGF on HUVECs and NIH 3T3 cells and promoted attachment of HUVECs, NIH 3T3 cells, AKR2B cells, and mink lung epithelial cells at 5–20 µg/ml (15). These latter effects occurred at 100- to 1000-fold higher concentrations than were needed for the stimulation of mitosis, chemotaxis, or ECM production by CTGF in other studies (10, 13, 14). Nonetheless, a role for CTGF in cell adhesion is further supported by the localization of CTGF in ECM (15) and by the ability of antisense CTGF to suppress growth and migration of BAECs in vitro (43). While 38-kDa CTGF appears to be proteolytically cleaved into low mass derivatives (11), the relationship (if any) between processing and activity remains undefined.

Although TGF-ß induces CTGF expression and CTGF mimics some of the effects of TGF-ß in fibroblasts, CTGF-independent pathways of TGF-ß action have been identified in these and other cell types. For example, TGF-ß is a potent inhibitor of the growth of mink lung epithelial cells, whereas CTGF is not (14). In addition, while CTGF and TGF-ß both stimulate DNA synthesis in monolayer cultures of NRK cells, only TGF-ß is able to stimulate anchorage-independent growth (AIG) of the same cells (14, 74). AIG induced by TGF-ß is antagonized by CTGF antibodies or antisense CTGF and is restored by addition of CTGF to TGF-ß-stimulated cells showing that CTGF is involved in the stimulation by TGF-ß of this process (74). Since CTGF was induced by TGF-ß in these cells, it was proposed that both CTGF-dependent and -independent pathways are involved in the stimulation of AIG by TGF-ß (74). The induction of CTGF mRNA and AIG by TGF-ß in NRK cells is inhibited by elevated cAMP levels (52). This effect was reversed by addition of CTGF, but not other growth factors, and was attributed to a CTGF-specific restriction point in late G₁ of TGF-ß-activated cells (52).

G. Mechanism of action of CTGF
In addition to contributing to TGF-ß-mediated AIG (see above), CTGF interacts synergistically with EGF, PDGF, IGF-I, or bFGF (10, 14, 56), suggesting that it activates distinct receptors and/or signaling pathways to those used by other growth factors. Although 38-kDa hCTGF was suggested to bind PDGF receptors (13, 75), this possibility was not supported using a mitogenic form of 10-kDa murine CTGF (39). Binding studies with ¹²⁵I-labeled 38-kDa rhCTGF have demonstrated the presence of specific high- and low-affinity binding sites and a 280-kDa cross-linked CTGF complex (76). Although there was no clear evidence for a signal-transducing receptor, these data are nonetheless potentially very exciting. Alternatively, CTGF may function primarily as a cell adhesion molecule that regulates cell function either directly through its association with the ECM or indirectly by synergizing with other growth factors (15).

	IV. Cyr61

Top Abstract I. Introduction II. The Modular Structure... III. Connective Tissue Growth... IV. Cyr61 V. Nov VI. Elm1/WISP-1 VII. Heparin-Induced CCN-Like... VIII. WISP-3 IX. Other CCN-Like Molecules X. Regulation of Cellular... XI. Biological Processes... XII. Perspectives and Future... References

 
A. Discovery of cyr61
Cyr61 (originally termed 3CH61) was described in 1985 as an immediate early gene in mouse Balb/c 3T3 cells that was induced by serum or PDGF (77). It was named cyr61 when shown to encode a cysteine-rich protein that contained 10% cysteine residues (78). The same gene, named ßIG-M1, was also discovered by Brunner et al. (31) in their studies of TGF-ß-inducible immediate early genes in mouse AKR-2B cells. The gene for cef-10, the chicken ortholog of cyr61, was described in 1989 as pp60^v-src-inducible immediate early gene in chicken embryo fibroblasts (CEFs) (79). cDNA for the human ortholog of cyr61 (hcyr61) was isolated from a 6-week-old human embryonic tissue cDNA library (80).

B. Structure of the cyr61 gene and protein
The murine cyr61 gene comprises five exons and four introns that span 3.1 kb (Table 2) and contains a serum-response element (SRE) (Fig. 4) in the 5'-region that mediates serum- or PDGF-induced gene transcription (6). Induction of cyr61 is similar to that of other immediate early genes that contain SREs, consistent with its coordinated expression with c-fos during G₀ to G₁ (81), although its transcriptional suppression is relatively inefficient (6). Although cyr61 expression is induced by TGF-ß (31), its promoter lacks the novel TGF-ß response element found in the promoter of CTGF (36) (Fig. 3). Human cyr61 maps to p22.3 on chromosome 1 (80, 82). The mouse cyr61 gene encodes a 379-residue protein, which, after signal peptide cleavage, yields a 355-residue protein containing 38 conserved cysteine residues (31, 78) (Fig. 1). The cysteine-free portion of cyr61 is about twice the length of that in other CCN paralogs (Fig. 1).

C. Cyr61 mRNA production
A single cyr61 transcript of 2.4 kb has been reported in mouse AKR2B cells, 3T3 fibroblasts, and PSA-1 teratocarcinoma cells (31, 77, 81, 83) and rat H19–7 embryonic neuronal hippocampal cells (84). A 1.8-kb cef-10 transcript is present in CEFs (79), and two cyr61 transcripts of 2.5 kb and 3.5–4 kb have been described in human tissues (80, 82). Although cef-10 mRNA is expressed principally in the adult lung (79), cyr61 is expressed in multiple adult tissues of the mouse and human such as heart, uterus, skeletal muscle, and lung (49, 78, 80). Transcript levels in other tissues are variable and likely due to differences in mRNA detection techniques as well as in the stage of development or differentiation of the source material. For example, human cyr61 mRNA expression is higher in the kidney of the fetus than that of the adult (80). Cyr61 mRNA is present at high levels in mouse embryos on days 9.5–14.5, whereas placental expression of cyr61 is highest on days 17.5–18.5 (83).

Cyr61 gene expression is stimulated by diverse molecular signals, which supports its role in a variety of biological processes. In mouse cells, cyr61 mRNA is rapidly induced by serum, PDGF, FGF, TGF-ß, phorbol ester, or cholera toxin (31, 77, 78, 81). Serum stimulation of Balb/c 3T3 cells resulted in a rapid increase and sustained high levels of cyr61 mRNA, which were stabilized by cyclohexamide (78). Similarly, human cyr61 mRNA is present in proliferating, but not quiescent, skin fibroblasts (49). In CEFs, transcription of cef-10 is rapidly enhanced by pp60^v-src or serum (79). In human osteoblasts, cyr61 expression is enhanced by EGF, PDGF, and interleukins-1ß, -2, and -6, as well as by 1,25-dihydroxyvitamin D₃, a mediator of osteoblast differentiation and function (85). In the rat uterus, cyr61 is estrogen-responsive and strongly induced by tamoxifen (86). In rat H19–7 cells, cyr61 is induced within 1 h by bFGF, a stimulator of neuronal differentiation in this cell type (84). This process appears to involve both mitogen-activated protein kinase-dependent and -independent pathways (84).

D. Cyr61 production and secretion
The 41-kDa mouse cyr61 protein is present at very low amounts in quiescent Balb/c 3T3 cells but reaches peak levels within 2 h of serum stimulation (78). Cyr61 is differentially distributed in the ECM, cellular fraction, or cell surface rather than the medium (71, 78). The extracellular fate and half-life of cyr61 synthesized in response to mitogenic stimulation were shown to be dependent upon the stage of the cell cycle and presence of binding moieties in the ECM and on cell surfaces such as heparin-like molecules from which cyr61 is eluted by 0.6–0.8 M NaCl (16, 49, 71).

E. Biological properties of cyr61
Recombinant murine cyr61 promotes dose-dependent attachment of HUVECs or NIH 3T3 cells to plastic surfaces coated with 3–30 µg/ml of the protein (16). Similar concentrations of cyr61 also stimulate dose-dependent chemotaxis of NIH 3T3 cells (16) and human microvascular endothelial cells (HMVEC) (87). These concentrations of cyr61 were not mitogenic for NIH 3T3 cells or HUVECs, although they did potentiate the mitogenic activity of bFGF for these cell types (16, 49). This latter effect was abolished by cyr61 antiserum and attributed to the displacement by cyr61 of bFGF from the ECM, thus increasing the effective concentration of soluble bFGF (16, 49). In mouse limb bud micromass cultures, cyr61 promoted expression of type II collagen, incorporation of sulfate, and the production of cartilage nodules (88). Moreover, cyr61 promoted the adhesion and aggregation of limb bud mesenchymal cells. Cyr61 was also a weak stimulator of DNA synthesis and proliferation of limb mesenchymal cells (88). Cyr61 was further shown to promote chondrogenesis in cell cultures seeded at subthreshold densities that do not normally undergo chondrogenic differentiation. These various responses were elicted with 0.3–5 µg/ml cyr61. Limb bud micromass cultures showed reduced levels of chondrogenesis when incubated with neutralizing cyr61 antisera, suggesting that endogenous cyr61 plays a normal physiological role in chrondrogenic differentiation (88). Finally, cyr61 has been shown to induce neovascularization in vivo (87), consistent with its stimulation of directed migration of HMVECs in vitro (87). Stable transfection of cyr61 cDNA into RF-1 gastric adenocarcinoma cells had no effect on in vitro growth rate, yet produced larger tumors in immunocompromized mouse hosts as compared with a vector control, a phenomenon that was attributed to cyr61-stimulated tumor vascularization (87).

F. Mechanism of action of cyr61
A mechanistic framework for the biological properties of cyr61 has been provided by the finding that cyr61 binds to integrin _vß₃, which represents the first (and only) molecularly defined receptor for any member of the CCN family (89). The interaction of _vß₃ with cyr61 may account for its promotion of chemotaxis and growth factor-mediated DNA synthesis as well as cell adhesion since integrins are known to modulate cell migration and growth factor signaling in other systems (90). As well as demonstrating a direct interaction between the two molecules in binding assays in vitro (89), cyr61-mediated adhesion and migration of cultured endothelial cells were specifically inhibited by the peptide RGDS and/or antiintegrin _vß₃ (87, 89). In addition to its integrin-binding property, cyr61 appears to be localized to its site of synthesis by associating with the ECM, possibly by binding to heparin-like molecules. This interaction may limit the extent of cyr61 diffusion so that its site of action is in close proximity to its site of synthesis (15).

	V. Nov

Top Abstract I. Introduction II. The Modular Structure... III. Connective Tissue Growth... IV. Cyr61 V. Nov VI. Elm1/WISP-1 VII. Heparin-Induced CCN-Like... VIII. WISP-3 IX. Other CCN-Like Molecules X. Regulation of Cellular... XI. Biological Processes... XII. Perspectives and Future... References

 
A. Discovery and structure of nov
Nov (nephroblastoma overexpressed) was first recognized as an overexpressed gene in nephroblastomas induced by myeloblastosis-associated virus type 1 (MAV-1) in day-old chicks (7, 91). Subsequently, the human (novH), mouse nov (novM), quail (novQ), and Xenopus (xnov) orthologs of chicken nov (novC) were isolated by hybridization screening (8, 33, 38, 92). The nov gene demonstrates a well conserved intron/exon structure that is similar to that of other paralogs in the CCN family (Table 2). In one chicken nephroblastoma, MAV-1 envelope and long-terminal repeat sequences integrated into the second exon of the novC gene at Cys⁶³ (7); the first putative initiation codon downstream of this integration site is Met⁹⁹. The novH gene maps to chromosome 8q24.1 and appears to contain at least three transcription initiation sites in the promoter region (38). NovM maps to chromosome 15 between D15 Mit153 and D15 Mit 183 (92, 93).

Putative transciption factor-binding sites are present in the 5'-untranslated region of several nov genes (8, 33, 38, 92) (Fig. 4). However, SREs and the TGF-ß response element are absent, suggesting that transcriptional regulation of nov is distinct from that of CTGF or cyr61. The activity of the novH promoter is repressed in the presence of the Wilms tumor suppressor gene WT-1 (94). While WT-1 binding sites are present in the novH promoter (Fig. 4), WT-1-mediated repression of novH transcription was experimentally attributed to other domains in the promoter (94).

The primary translational products of nov orthologs are predicted to comprise between 343 and 357 amino acids and to contain 38 conserved cysteine residues (Fig. 1). The signal peptide in novH appears to comprise 27 residues (8). In CEFs, an immunoreactive 46-kDa novC protein was localized to the ECM (93), although the underlying mechanisms and functional significance remain unclear. NovH produced in transfected Madin Darby canine kidney cells is a glycosylated protein that is secreted about 1.5 h after synthesis and requires 0.5 M NaCl for elution from heparin (20). Secreted novH has a half-life of more than 18 h, which is more than 10 times that of intracellular novH (20). Levels of the endogenous 46-kDa novH protein are repressed in 293 kidney cells that constitutively express WT1 (94).

B. Nov mRNA production
A single 2.2-kb novC transcript is usually detected in MAV1-induced nephroblastomas (7). An exception was a tumor in which MAV1 integrated within the novC gene itself, resulting in the production of a 2.0-kb transcript (7). The 2.2-kb novC mRNA transcript was detected at high levels in the brain and heart of day 18 chick embryos, with weak signals in muscle and intestine and nondetectable levels in liver, lung, and yolk sac. In the adult, 2.2-kb novC mRNA was expressed at high levels in brain and lung, at low levels in the spleen, and was nondetectable in heart, muscle, and liver (7). Quiescent cultures of CEFs contain a 2.2-kb novC transcript that is down-regulated during proliferation, mitogenic stimulation, or activation of viral oncogenes (95). NovH is normally expressed as a 2.5-kb transcript in human glioma cell lines and Wilms tumors, although an additional novH mRNA of 1.2 kb was observed in a single Wilms tumor (8, 37). Unlike novC, this smaller transcript was not the result of rearrangement of the novH gene (8).

There is little, if any, experimental evidence supporting a role for nov as an immediate early gene. As explained above, there are no motifs in the nov gene promoter that resemble either a SRE or a TGF-ß response element. Serum stimulation of kidney-derived 293 cells or CEFs failed to increase nov mRNA levels within 90 min, even though the genes for CTGF and jun were rapidly activated (93). Moreover, in normal CEFs, nov was expressed in quiescent cells but down-regulated in proliferating cells (95). In contrast to cef-10, expression of p60^v-src in CEFs resulted in transcriptional down-regulation of the novC gene (95). NovC expression was also repressed by the viral oncogenes v-erbB, v-mil, and v-crk (95). In addition, mRNA destabilization sequences in the 3'-region are absent from most nov orthologs (except xnov), suggesting that nov RNA degradation is regulated differently from that of CTGF, cyr61, and other immediate early genes.

C. Biological properties of nov
Overexpression of full-length novC cDNA was shown to inhibit the growth of CEFs grown in soft agar (7). In contrast, transformation of CEFs occurred after overexpression of an N-terminally truncated form of novC that initiated at Met⁹⁹ and was thus comparable to that expressed after integration of MAV-1 into the novC gene (Fig. 3) (7). This truncated form of nov also induced morphological transformation of rat embryo fibroblasts (8). Within the CCN family, the ability to function as a protooncogene is thus far unique to nov. The dramatic biological differences between the normal and truncated nov genes have led to suggestions that nov is a growth suppressor gene, truncation of which results in oncogenic activation. These interpretations are consistent with the low levels of nov mRNA in mitogen-stimulated cells and the high levels of nov mRNA in resting cells (93, 95). Although the truncated nov protein lacked the signal peptide and the IGF-binding motif in module 1, it is not known how the loss of one or both of these domains contributes (if at all) to its transforming properties.

	VI. Elm1/WISP-1

Top Abstract I. Introduction II. The Modular Structure... III. Connective Tissue Growth... IV. Cyr61 V. Nov VI. Elm1/WISP-1 VII. Heparin-Induced CCN-Like... VIII. WISP-3 IX. Other CCN-Like Molecules X. Regulation of Cellular... XI. Biological Processes... XII. Perspectives and Future... References

 
Elm1 (expressed in low-metastatic type 1 cells) was initially identified by mRNA differential display as being expressed in low, but not in high, metastatic murine melanoma cells (96). The elm1 protein was subsequently predicted to comprise 367 amino acids and to conform to the CCN modular structure (97) (Figs. 1 and 3; Table 1). Elm1 transcripts of 1.8 and 5.0 kb were present in all mouse tissues analyzed (97). The 5.0-kb elm1 transcript was induced within 3 h of serum stimulation of quiescent Balb/c 3T3 cells, unlike cyr61 which was induced within 30 min in the same cells (97). Elm1 maps to mouse chromosome 15 between D15 Mit17 and D15 Mit3. Southern blotting demonstrated that the elm1 gene is also present in human, monkey, rat, mouse, dog, and cow (97). Transfection of elm1 into highly tumorigenic and metastatic K-1735 M-2 cells resulted in a decreased tumor growth in vivo and reduction in the frequency of lung metastatic colonies (97). In vitro, transfected cells that expressed relatively high amounts of elm1 grew somewhat slower and to lower saturation density as compared with control cells (97). Elm1 did not exhibit mitogenic activity for NIH3T3 cells, either alone or in combination with bFGF. The growth-suppressive properties of elm1 have been tentatively linked to module 3 (97). Recently, WISP-1 was identified as the human ortholog of elm1 and shown to be a component of the Wnt-signaling pathway in transformed cells (24). WISP-1 maps to human chromosome 8q24.1–8q24.3 and exhibits tissue-specific patterns of expression (24).

	VII. Heparin-Induced CCN-Like Protein (HICP)/rCop-1/CTGF-3/WISP-2

Top Abstract I. Introduction II. The Modular Structure... III. Connective Tissue Growth... IV. Cyr61 V. Nov VI. Elm1/WISP-1 VII. Heparin-Induced CCN-Like... VIII. WISP-3 IX. Other CCN-Like Molecules X. Regulation of Cellular... XI. Biological Processes... XII. Perspectives and Future... References

 
A novel 1.8-kb mRNA was identified by subtractive hybridization screening of heparin-treated and nontreated rat VSMCs and shown to encode a novel member of the CCN family that was termed HICP (98). Independent studies resulted in the isolation of cDNA for the same gene, termed rCop-1, when shown to be expressed in normal rat embryo fibroblasts or mouse 3T3 cells, but not in their transformed derivatives (99). The mouse ortholog, WISP-2, was recently identified as a unique gene that was up-regulated in Wnt-1-transformed mouse mammary epithelial cells in vitro (24). These molecules, together with the human ortholog, CTGF-3 (100), demonstrate an overall amino acid homology of 30% with hCTGF and conservation of all of the cysteine residues in modules 1–3 (Fig. 1). However, all of these proteins are C-terminally truncated as compared with other CCN paralogs, resulting in a total absence of module 4 (Figs. 1 and 3). While HICP/CTGF-3/WISP-2 transcripts were present in multiple tissues (24, 98, 100), rCop-1 mRNA was not detected in any of the major organs of the mouse embryo and adult rat (99). Although this discrepancy requires further investigation, expression of rCop-1 has been linked to aging and senescence (99). HICP expression patterns, similar to those of growth arrest genes, occur at high levels in quiescent or heparin-arrested VSMCs and at very low levels during proliferation (98). Somewhat different expression patterns occur for rCop-1 in cycling mouse 3T3 cells in which rCop-1 mRNA is absent during quiescence or immediately after serum stimulation and appears only during late S phase (99). Unlike normal cells, transfection of rCop-1 into transformed cells suppressed their cell growth; this was attributed to cell death rather than growth arrest (99). In transfected cells, rCop-1 is localized primarily intracellularly and to the cell surface, but not in conditioned medium or ECM. It has been speculated (99) that the unique subcellular localization and expression pattern of rCop-1/HICP is a reflection of its role in negative growth regulation, the structural basis of which may lie in the absence of module 4. Potential tumor-suppressive properties are further suggested by the finding that WISP-2 is underexpressed in human colon tumors, unlike WISP-1 and -3, which are overexpressed (24).

	VIII. WISP-3

Top Abstract I. Introduction II. The Modular Structure... III. Connective Tissue Growth... IV. Cyr61 V. Nov VI. Elm1/WISP-1 VII. Heparin-Induced CCN-Like... VIII. WISP-3 IX. Other CCN-Like Molecules X. Regulation of Cellular... XI. Biological Processes... XII. Perspectives and Future... References

 
Human WISP-3, a 354-residue protein containing 36 cysteine residues, was identified by screening expressed sequence tag databases with WISP-1 (24). The secreted WISP-3 protein is predicted to contain 34 of the 38 conserved cysteine residues found in most other CCN proteins since it lacks 4 of the cysteines normally present in module 2. It is possible that this structural difference results in unique properties and functions, although this aspect of WISP-3 biology has not yet been explored (24).

	IX. Other CCN-Like Molecules

Top Abstract I. Introduction II. The Modular Structure... III. Connective Tissue Growth... IV. Cyr61 V. Nov VI. Elm1/WISP-1 VII. Heparin-Induced CCN-Like... VIII. WISP-3 IX. Other CCN-Like Molecules X. Regulation of Cellular... XI. Biological Processes... XII. Perspectives and Future... References

 
Twisted gastrulation (tsg) and short gastrulation (sog), which are involved in dorsoventral patterning in Drosophila embryos (101, 102), contain sequences that resemble, respectively, modules 1 and 2. As with CTGF and cyr61, tsg is eluted from heparin by 0.6–0.8 M NaCl, and the binding of tsg by somatic cells may involve a heparin-like coreceptor (103). The cytoskeletal rearrangements that take place in dorsal amnioserosa cells in the presence of tsg have been likened to those in endothelial cells during CTGF-induced chemotaxis (101). Sog, like CTGF, may be susceptible to proteolytic cleavage, yielding diffusible bioactive fragments (102). The cDNA of "small CCN-like growth factor" (SCGF) was recently isolated from an 9-week human embryo library and shown to encode a 206-residue protein that had limited structural similarity to the C-terminal region of CCN family members, although cysteine residues occur at half the expected frequency and are poorly conserved (104). Overall, tsg, sog, and SCGF exhibit very weak alignment with the CCN family and are not genuine paralogs.

	X. Regulation of Cellular Functions by the CCN Family

Top Abstract I. Introduction II. The Modular Structure... III. Connective Tissue Growth... IV. Cyr61 V. Nov VI. Elm1/WISP-1 VII. Heparin-Induced CCN-Like... VIII. WISP-3 IX. Other CCN-Like Molecules X. Regulation of Cellular... XI. Biological Processes... XII. Perspectives and Future... References

 
A. Cell cycle control
Members of the CCN family are products either of immediate early genes (CTGF, cyr61) or of putative growth arrest/suppression genes (nov, elm1, HICP). The expression of immediate early genes represents the earliest genomic response to growth factors and is likely to initiate the program leading to cell replication. Since they encode a diverse array of regulatory molecules, the induction of immediate early genes represents a central component in the proliferative response to mitogenic stimuli (105). Since CTGF and cyr61 are transcriptionally activated by TGF-ß, PDGF, EGF, FGF, TPA, and cholera toxin, it is likely that the cellular responses to these diverse mitogenic stimuli are controlled, at least partly, by CTGF and cyr61. On the other hand, the expression of growth arrest-specific genes, DNA damage-inducible genes, and the MyD growth-arrest genes are associated with the negative regulation of cell growth (106). Growth arrest genes have been implicated in cellular differentiation, embryonic development, and apoptosis (107, 108, 109), and it is possible that these types of processes are similarly regulated by nov, elm1, and rCop-1.

In addition, each CCN family member appears to further regulate the cell cycle through their 1) direct mitogenic action (e.g., CTGF); 2) potentiation of the mitogenic activity of other growth factors (e.g., CTGF, cyr61); 3) binding of IGFs (e.g., CTGF); 4) regulation of ECM synthesis (e.g., CTGF, cyr61); 5) regulation of cell attachment and migration (e.g., CTGF, cyr61, elm1); 6) interactions with the ECM (e.g., CTGF, cyr61, nov); 7) oncogenic properties (e.g., nov); and 8) regulation of a cell cycle restriction point (e.g., CTGF). This broad spectrum of biological properties, many of which are interacting and interdependent, demonstrate the complexity by which the cell cycle is likely regulated by the CCN family.

B. Cell adhesion and migration
In vitro studies have shown that CTGF or cyr61 are chemotactic and promote cell adhesion (13, 15, 16, 87). Elm1 is expressed in low, but not high, metastatic cell lines and, when transfected into high-metastatic cell lines, causes them to exhibit slower rates of tumor growth and decreased incidence of metastasis (97). Truncated nov stimulates AIG and colony formation in CEFs (7). Collectively, these data demonstrate broad effects on cell adhesion and locomotion, which likely reflects the ability of CCN family members to regulate the composition of the ECM, the net balance between its synthesis and degradation, as well as to bind directly to ECM components such as integrins and heparan sulfate proteoglycans (see below).

C. ECM production
Tissue formation and cell proliferation and differentiation are dependent upon interactions between cells and the ECM. In processes such as tissue remodeling and malignancy, ECM components are degraded by proteases such as collagenase, plasmin, and matrix metalloproteases, and the ability of cells to interact with the ECM is altered or abolished. TGF-ß plays a central role in ECM production since it stimulates synthesis of ECM proteins such as collagen, fibronectin, laminin, elastin, glycosaminoglycans, and other glycoproteins as well as synthesis of inhibitors of ECM degradation such as plasminogen activator inhibitor and tissue inhibitors of metalloproteases (110). In view of the TGF-ß-inducibility of CTGF and cyr61 gene expression, these molecules may promote ECM deposition by stimulating synthesis of ECM components and inhibiting their degradation. While detailed studies of the regulation of ECM-degrading enzymes and their inhibitors by the CCN family have yet to be reported, initial studies have shown that 38-kDa CTGF enhances fibronectin, type IV collagen, and integrin production (14) and that cyr61 stimulates type II collagen production, albeit in a specialized cell type (88). As discussed below, numerous fibrotic disorders exhibit substantial ECM involvement and expression of CTGF. In addition, there is a striking appearance of CTGF in uterine stromal cells that are undergoing decidualization (47), a highly regulated process involving increased mitosis and synthesis and deposition of ECM molecules such as desmin, laminin, and fibronectin (111). ECM production and remodeling is also a critical component of development and differentiation during embryogenesis (112). Collectively, these data suggest that CTGF may exert diverse and important functions in normal physiology and pathological states via its net induction of ECM protein deposition and content.

	XI. Biological Processes Involving the CCN Family

Top Abstract I. Introduction II. The Modular Structure... III. Connective Tissue Growth... IV. Cyr61 V. Nov VI. Elm1/WISP-1 VII. Heparin-Induced CCN-Like... VIII. WISP-3 IX. Other CCN-Like Molecules X. Regulation of Cellular... XI. Biological Processes... XII. Perspectives and Future... References

 
A. Development and differentiation
Immunohistochemical studies showed that CTGF is present in the mouse as early as embryonic days 4.5–6.5, at which time it is most abundant in the embryonic endoderm and mesoderm (47). At later stages of gestation (days 14–18), CTGF and cyr61 are both present in various tissues and organs including the cardiovascular and pulmonary systems as well as the skin and placenta (15). Secretory structures such as kidney tubules and salivary, mucous, and sebaceous glands are positive for CTGF, but not cyr61, whereas the nervous and skeletal systems are positive for cyr61, but not CTGF (15). This differential distribution may be indicative of unique roles for each factor. A role for nov during Xenopus development is suggested by the production of its mRNA in the oocyte at stages I, III, and IV and in the embryo at all developmental stages between egg and tadpole (33). In chickens, novC is expressed in kidney, heart, and muscle of the embryo but not of the adult (7). Other organs (e.g., brain) demonstrate novC expression in both embryonic and adult life, whereas expression of novC in the lung occurs in the adult but not the embryo (7).

Evidence to date supports a role for cyr61 and CTGF in chondrogenesis. The cyr61 protein is present in cartilage and bone in day 13–18 mouse embryos (15, 88), and cyr61 mRNA is present in a variety of developing cartilaginous structures in day 8.5–14.5 embryos (83). A role for cyr61 in normal growth and development of the cartilaginous skeleton is further supported by the ability of cyr61 to stimulate collagen synthesis, sulfate incorporation, and formation of cartilage nodules in mouse limb bud micromass cultures, and adhesion, aggregation, and growth of limb bud mesenchymal cells (88). In the case of CTGF, its mRNA is present in hypertrophic chondroctyes from day 17 mouse embryos, neonatal rabbit growth cartilage tissue and cultured cells, and human chondrocytic cell lines, but not in human osteosarcoma or mouse osteocytic cell lines (48). In cultured rabbit growth cartilage cells, peak levels of CTGF occurred during the early hypertrophic stage. CTGF mRNA levels in these cells were enhanced by treatment with TGF-ß or bone morphogenic protein-2 (48). In addition to its potential role in chondrogenesis, cyr61 may also be involved in hippocampal differentiation, since it is induced in H19–7 cells by bFGF, which stimulates their differentiation into nonproliferative neuronal cells (84).

Several lines of evidence support a role for nov nephrogenesis. First, novC expression occurs in normal avian kidney cells of the embryo and neonate, yet is barely detectable in adult kidney cells (7). Second, ureteric buds, which give rise to the kidney collecting ducts, fail to demonstrate outgrowth in WT-1-null mice; as discussed above, novH promoter activity is repressed by WT-1, and a reciprocal quantitative gradient of novH and WT1 expression occurs during normal human nephrogenesis (see Ref. 94). Third, novH is localized to differentiating glomerular podocytes where it accumulates and may regulate podocyte structure or function during both pre- and postnatal life (20). Since the composition of the ECM changes during the earliest stages of metanephric induction (113) and ECM components regulate uteric bud morphogenesis, nov may also influence nephrogenesis by binding to the ECM (93).

B. Female reproductive tract function
The actions of ovarian steroids are mediated, in part, by polypeptide growth factors, which may also contribute to the uterine-embryo signaling dialogue that initiates implantation and stimulates embryonic development (114, 115, 116, 117, 118, 119, 120). Also, some growth factors are secreted into the uterine lumen or localized at the implantation site, where they may stimulate development of the embryo and placental membranes (114, 121). CTGF is present in uterine fluids of the pig and mouse, as well the uterus of the pig, mouse and human (10, 11, 13, 47, 56, 122) suggesting that it is involved in the regulation of uterine function. In uterine fluids, the levels of low mass CTGFs and CTGF proteases are highly correlated with each other and show cyclic variations during the estrous cycle (11). Moreover, there are clear differences in the levels of CTGF and CTGF protease(s) in uterine fluids at equivalent stages of the estrous cycle and early pregnancy suggesting that CTGF production or action is modified in the presence of the embryo (11, 56). A relationship between CTGF expression and TGF-ß action during the periimplantation period of the pig is supported by the presence of both molecules at this time in uterine and/or embryonic tissues (11, 123, 124, 125). In addition, a direct role of CTGF on uterine cells is indicated by its stimulation of DNA synthesis in pig stromal cells in vitro (56).

In mice and women, CTGF is localized primarily to uterine luminal and glandular epithelial cells during the estrous cycle and during the first few days of pregnancy (47, 122). On the day of implantation in mice, epithelial staining for CTGF is strongly reduced and is followed over the next 2 days by profound staining of decidualizing endometrial stromal cells, suggesting that CTGF contributes to the decidualization process or is produced as a result of it (47). Decidualization is a highly regulated differentiation process involving increased vascular permeability, DNA synthesis, and synthesis and deposition of ECM molecules (111, 126, 127, 128). The distribution of uterine CTGF during early pregnancy in the mouse does not entirely correlate with TGF-ß and its receptors, suggesting that TGF-ß-independent synthesis of CTGF may be operative (47). Alternative mechanisms may involve glucocorticoids or estrogen, which stimulate CTGF and cyr61 gene transcription, respectively (55, 86). The nonconcordance of CTGF and TGF-ß may also be related to the initial production of TGF-ß in a latent form (129). Whatever the explanation, these data support a role for uterine CTGF at the time of implantation.

C. Angiogenesis
Angiogenesis, the formation of new capillaries from preexisting blood vessels, occurs in processes such as tumor growth, diabetic retinopathy, wound healing, and placental vascularization. It is a complex process that involves degradation of the capillary basement membrane, migration and proliferation of endothelial cells, and tube formation; it is regulated by many factors (130), including cyr61, which promotes directed migration of HMVECs via _vß₃-integrin and induces neovascularization in the rat cornea (87). Expression patterns of cyr61 support its contribution to angiogenesis in the embryo, placenta, hypertrophic cartilage, wounds, and tumors (83, 87). Since cell migration is the only component of angiogenesis to be directly stimulated by cyr61, indirect effects of cyr61, such as release and action of the angiogenic factor bFGF from ECM (16, 49), may contribute to the overall process of neovascularization. These questions and the role of other CCN family members in angiogenesis will require further study.

D. Wound repair
Many studies have established that growth factors, including TGF-ß, are involved in wound healing (see Ref. 131). Collectively, CTGF and cyr61 exhibit numerous biological properties that are of potential importance in the wound healing response, including stimulation of cell proliferation, cell adhesion, chemotaxis, angiogenesis, production of ECM components, and augmentation of bFGF activity. Early studies showed that cyr61 gene expression was induced during the first hour of liver regeneration in the mouse after partial hepatectomy (132). These results are consistent with the role of cyr61 as an immediate early gene and demonstrate that cyr61 is transcriptionally activated as a response to injury. In tissue from within subcutaneously implanted Schilling chambers in rats (133), CTGF expression peaked on day 9 of injury after peak expression of TGF-ß on day 3 (41). The coordinate expression of the two factors was interpreted as a component of a growth factor cascade in which TGF-ß initiated regeneration and repair and stimulated the production of CTGF, which was required later in the wound healing process (41). Somewhat different kinetics of gene transcription have been reported in full thickness wounds in which CTGF and TGF-ß both exhibited peak expression 12–24 h after injury (55, 134). Although dexamethasone was shown to stimulate CTGF gene expression in uninjured tissue, it did not affect CTGF mRNA levels in wounded tissues and was attributed to the presence of TNF-, which inhibits CTGF gene expression (35, 55). Finally, scrape wounding BSC-1 renal epithelial cells resulted in stimulation of CTGF gene expression, which reached peak levels 4 h after injury (46). CTGF was induced both in cells at the wound and some distance from it, suggesting that an inductive signal was communicated to distant cells from the initial wound site, although there were no changes in expression levels of TGF-ß, PDGF, or bFGF (46).

E. Fibrotic disorders
Many fibrotic disorders are typified by excessive connective tissue and ECM formation and exhibit marked overexpression of TGF-ß, which is fibrogenic and strongly linked to the pathogenesis of these diseases (135, 136, 137). In view of the biological activities of CTGF and the link between CTGF production and TGF-ß action, studies have recently been initiated to examine the role of CTGF in fibrotic disorders of the skin, kidney, lung, and blood vessels. Subcutaneous injection of TGF-ß into neonatal mice, which causes a rapid increase in the amount of granulation tissue comprising connective tissue cells and abundant ECM, was shown to result in enhanced levels of CTGF mRNA in connective tissue fibroblasts but not in epithelial cells or endothelial cells (14, 72). Injection of 38-kDa rhCTGF caused a very similar fibrotic reaction as TGF-ß in terms of histological findings, time course, and area affected (14). This fibrotic response was specific to TGF-ß and CTGF and not mimicked by EGF, PDGF, or bFGF (14, 72).

Overexpression of CTGF mRNA occurs in a variety of fibrotic skin disorders including 1) systemic sclerosis in which CTGF expression is temporally associated with the sclerotic phase and is highest in the fibroblasts of the deep dermis (42); 2) localized sclerosis in which CTGF-positive fibroblasts are scattered throughout the lesion (138); 3) keloids in which CTGF-positive fibroblasts are present throughout the lesion but concentrated in the expanding peripheral regions (138); and 4) scar tissue, eosinic fasciitis, nodular fasciitis, and Dupuytren’s contracture in which CTGF was partially expressed in some of the fibroblasts (138).

In kidney fibrosis, CTGF expression is increased in inflammatory glomerular and tubulointerstitial lesions, as compared with normal kidney or noninflammatory glomerular lesions (139). Extracapillary proliferative lesions, capsular adhesions, and periglomerular fibrosis were characterized by a pronounced increase in epithelial expression of CTGF. CTGF expression was also somewhat increased in mesangial proliferative lesions of diabetic and IgA nephropathies (139). In a rat model of glomerulonephritis, CTGF mRNA was up-regulated in parietal epithelial cells and podocytes, where it was expressed in areas of crescentic extracapillary proliferation, periglomerular fibrosis, and in interstitial foci (140). Cultured mesangial cells and podocytes demonstrated enhanced CTGF mRNA in response to TGF-ß (140) or under hyperglycemic conditions (141), the latter of which causes ECM deposition and induction of TGF-ß1 (142).

In bleomycin-induced fibroproliferative lung disease in mice, which is characterized by up-regulation of TGF-ß mRNA (143, 144), bleomycin treatment of sensitive mice resulted in a 2- to 3-fold increase in lung CTGF mRNA levels and collagen synthesis as compared with resistant mice (34). CTGF is present at higher levels in bronchoalveolar lavage fluid from patients with fibrosing alveolitis as compared with normal individuals, and lung fibroblasts from patients with scleroderma-associated fibrotic lung produce more CTGF in response to TGF-ß than control cells (145). An autocrine role for CTGF in lung growth is supported by the expression of CTGF mRNA in murine lung (5) and in cultures of human or mouse lung mesenchymal cells in which it is rapidly induced by TGF-ß (34) and its stimulation of mitosis and collagen synthesis in fibroblasts (73, 145).

CTGF mRNA is expressed in atherosclerotic vessels at 50- to 100-fold the level of that in normal arteries (44). In advanced atherosclerotic lesions, CTGF was localized to VSMCs and endothelial cells primarily at sites of ECM accumulation and fibrosis, including the shoulders of fibrous caps (44, 139, 146). VSMCs respond mitogenically to CTGF (10) and produce CTGF mRNA in response to TGF-ß (44, 146), consistent with the presence of a CTGF autocrine loop that is initiated by TGF-ß. Although the potential involvement of CTGF in stimulating intimal thickening is highly deleterious, it has been argued that CTGF-stimulated ECM production may actually stabilize the fibrous cap and reduce the chance of plaque rupture (146). Finally, the association between HICP expression and growth arrest in VSMCs (98) suggests that attenuation of HICP may contribute to hyperplastic VSMC diseases such as athero- and arteriosclerosis.

The profibrotic properties of CTGF suggest that it is an attractive therapeutic target in a variety of fibrotic disorders, especially as it operates downstream of TGF-ß (53, 136). Intervention at the level of CTGF would still allow certain beneficial non-CTGF-dependent effects of TGF-ß (e.g., antiinflammatory) to persist while negating its fibrogenic action. Over the next few years we can expect to see a careful evaluation of all components of the CTGF pathway—its gene, mRNA, transcription factors, protein, receptor, and second messengers—with the goal of finding optimal molecular targets for the control of CTGF-mediated fibrosis.

F. Inflammation
In addition to inflammatory kidney disease (see above), CTGF is overexpressed in inflammatory bowel diseases (IBD) such as Crohn’s disease and ulcerative colitis (147). IBD demonstrated high levels of CTGF, TGF-ß, collagen type I, fibronectin, and integrin 5. CTGF was overexpressed in regions of inflammation and in noninflamed regions that were stenosed and was present in the vicinity of TGF-ß-producing cells (147). CTGF was thus proposed to promote mesenchymal tissue repair and remodeling after the acute inflammatory phase and to stimulate chronic matrix deposition leading to fibrosis and stenosis (147).

G. Tumor growth
Several lines of evidence support a role for CCN molecules in tumorigenesis, the most compelling of which is the oncogenic property of N-terminally truncated nov (7). The tumorigenicity of a gastric adenocarcinoma cell line was increased when transfected with cyr61, the angiogenic properties of which support its role in tumor growth and vascularization (87). Consistent with its profibrotic properties, CTGF is overexpressed in breast cancer, pancreatic cancer, and melanomas exhibiting significant involvement of connective tissue cells (desmoplasia) (54, 148, 149). Similarly, WISP-1 and WISP-2 are strongly expressed in the fibrovascular stroma of breast tumors from Wnt-1 transgenic mice (24). CTGF is expressed in sarcoma and chondrosarcoma cells (39, 48), while cyr61 is expressed in rhabdomyosarcoma and cell lines derived from malignant melanoma, breast and colon adenocarcinoma, and bladder papilloma (87, 150). Tumors of the nervous system express CTGF, nov, and cyr61 in a complex and mainly noncorrelative pattern (37, 82).

Elevated expression of novC mRNA was a consistent finding in all MAV1- and MAV2-induced avian nephroblastomas (7, 93). While novC was expressed most highly in differentiated tumors (7), its levels are more correlated with tumor age than histological status (93). In Wilms tumors, novH is mainly overexpressed in tumors of predominantly stromal origin (8) and is positively correlated with heterotypic muscle differentiation (20). Nov expression is inversely correlated with the levels of WT-1 mRNA in some (8, 93), but not all (20), studies. Since WT-1 and several viral oncogenes repress nov expression in vitro (94, 95), these variable results demonstrate that the relationship between nov expression and tumorigenesis is complex and that nov transcription mechanisms in both normal and tumor cells requires further study. Despite its apparent role as a growth arrest gene in transfected cells (7), the potential role of nov in growth arrest may be cell type specific or secondary to other biological functions in tumors, thereby accounting for its overexpression in some nephroblastomas (7, 93). Nonetheless, the absence of the IGF binding motif in oncogenic nov and the coexpression of nov and IGF-II in some avian renal tumors and Wilms tumors suggest a possible link between nov and IGF in nephroblastoma (93).

The inverse relationship between nov expression and phenotype of some nephroblastomas is shared with other CCN family members in other tumors. For example, elm1 expression is inversely correlated with the incidence of metastasis and growth of melanoma cells (96, 97). In addition, inverse correlations have been reported between malignant phenotype and the level of CTGF expression in fibroblast and endothelial cell tumors (151), and the level of cyr61 expression in neuroblastoma (83) and prostate cancer (152). On the other hand, rCop-1 is underexpressed in transformed cells and is a negative regulator of the growth of transformed, but not normal, cells (99). Colon tumors demonstrate underexpression of WISP-2 and overexpression of WISP-1 and -3 while in breast tumor models both WISP-1 and -2 are involved in the Wnt signaling pathway that leads to cell transformation (24). Collectively, the individual CCN family members appear to be over-, under-, or randomly expressed in a variety of tumors, and it is currently impossible to establish a unifying hypothesis for the CCN family in tumor growth.

	XII. Perspectives and Future Directions

Top Abstract I. Introduction II. The Modular Structure... III. Connective Tissue Growth... IV. Cyr61 V. Nov VI. Elm1/WISP-1 VII. Heparin-Induced CCN-Like... VIII. WISP-3 IX. Other CCN-Like Molecules X. Regulation of Cellular... XI. Biological Processes... XII. Perspectives and Future... References

 
It is clear that the initial classification of CCN proteins as immediate early gene products or growth factors is not universally true for all members of this family. As the family has grown, so has its spectrum of biological properties. In fact, the range of activities within the CCN family is now so broad that their classification on a functional basis is difficult, if not impossible. While they have been categorized by several investigators as ECM or extracellular signaling molecules, this is certainly not a distinguishing feature of the CCN family nor particularly unexpected for modular proteins. Moreover, the signaling events themselves are poorly defined at best. In view of their modular structure and presumed ability to interact with a diverse array of proteins in the pericellular environment, CCN proteins can be expected to demonstrate complex regulation in time and space. As with other bioactive proteins such as growth factors (153) and modular proteins such as TSP-1 (154), interactions with binding proteins are predicted to substantially impact the net biological properties and bioavailability of a given CCN member. While this aspect of CCN biology has not yet been systematically studied, it might help to explain some of the discrepancies and apparent contradictions that have been reported between various laboratories regarding the localization and activities of individual proteins. The likely involvement of the CCN family in complex protein-protein interactions further indicates that a careful analysis of the modules, individually and collectively, is key to understanding the relative properties of a given ortholog and its processed forms. Informative data will likely come from powerful molecular strategies such as the yeast two-hybrid system which was recently utilized to identify the binding of fibulin 1C to novH (155). In addition, the very complex molecular configuration of the CCN family highlights the importance of verifying that recombinant forms of the family members are appropriately folded and faithfully reproduce the biological properties of their native counterparts; this is no small challenge and has been largely overlooked to date. Based on other modular proteins (154), a much broader perspective is needed when considering the activities of CCN proteins, the biological processes in which they act, and the mechanisms involved.

Many additional questions remain unanswered. Are there other members of the CCN family? Do CCN proteins physiologically bind IGFs and, if so, what are the consequences? Are there unique signal-transducing receptors for CCN proteins? What signaling pathways are activated when CCN proteins bind to cell surfaces? What are the identities of CTGF proteases and how are they regulated? What are the biological consequences of the absence of module 4 in rCop-1 and the absence of the cysteine residues in module 2 of WISP-3? Why is nov overexpressed in some nephroblastomas if it is a growth arrest gene? How does cyr61 promote angiogenesis? What other disease states involve the CCN family? What is the relationship between the CCN family and endocrine function? With recent interest in the field, rapid progress in answering some of these questions can be expected in the near future.

In conclusion, the CCN family comprises highly related modular proteins that have wide ranging properties that impact cellular functions such as growth, differentiation, adhesion, and locomotion. The biological properties and actions of CCN proteins are probably complex and likely reflect a dynamic equilibrium of their constituent modules with soluble, cell surface, or ECM-binding proteins. Since these conditions are hard to define molecularly and difficult, if not impossible, to reproduce in vitro, the challenge ahead is to understand the biological roles of the CCN family in normal and pathological processes as a function of the composition of the pericellular environment in vivo.

	Acknowledgments

 
I extend my sincere thanks to Cecile Martinerie, Lester Lau, David Abraham, Sabine Werner, and Christopher Wenger for providing me with preprints and abstracts of their recent work. I am especially indebted to John Castellot for furnishing me with unpublished data on HICP. I am grateful to Gulnar Surveyor for reviewing the manuscript and thank Amy Wilson and DeAnna Ball for help with artwork.

	Footnotes

 
Address reprint requests to: D. R. Brigstock, Ph.D., Wexner Institute for Pediatric Research, Children’s Hospital, 700 Children’s Drive, Columbus, Ohio 43205 USA.

¹ This work was supported by NIH Grant HD-30334, US Department of Agriculture Grant 9803693, and FibroGen, Inc. (in which D.R.B. has an equity interest).

	References

Top Abstract I. Introduction II. The Modular Structure... III. Connective Tissue Growth... IV. Cyr61 V. Nov VI. Elm1/WISP-1 VII. Heparin-Induced CCN-Like... VIII. WISP-3 IX. Other CCN-Like Molecules X. Regulation of Cellular... XI. Biological Processes... XII. Perspectives and Future... References

 

1. Bork P 1993 The modular architecture of a new family of growth regulators related to connective tissue growth factor. FEBS Lett 327:125–130[CrossRef][Medline]
2. Bork P 1992 Mobile modules and motifs. Curr Opin Struct Biol 2:413–421[CrossRef]
3. Bork P 1991 Shuffled domains in extracellular proteins. FEBS Lett 286:47–54[CrossRef][Medline]
4. Patthy L 1991 Modular exchange principles in proteins. Curr Opin Struct Biol 1:351–361
5. Ryseck R-P, Macdonald-Bravo H, Mattéi M-G, Bravo R 1991 Structure, mapping and expression of fisp-12, a growth-factor-inducible gene encoding a secreted cysteine-rich protein. Cell Growth Differ 2:225–233[Abstract]
6. Latinkic BV, O’Brien TP, Lau LF 1991 Promoter function and structure of the growth factor-inducible immediate early gene cyr61. Nucleic Acids Res 19:3261–3267[Abstract/Free Full Text]
7. Joliot V, Martinerie C, Dambrine G, Plassiart G, Brisac M, Crochet J, Perbal B 1992 Proviral rearrangements and overexpression of a new cellular gene (nov) in myeloblastosis-associated virus Type 1-induced nephroblastomas. Mol Cell Biol 12:10–21[Abstract/Free Full Text]
8. Martinerie C, Huff V, Joubert I, Badzioch M, Saunders G, Strong L, Perbal B 1994 Structural analysis of the human nov proto-oncogene and expression in Wilms tumors. Oncogene 9:2279–2732
9. Patthy L 1987 Intron-dependent evolution: preferred types of exons and introns. FEBS Lett 214:1–7[CrossRef][Medline]
10. Brigstock DR, Steffen CL, Kim GY, Vegunta RK, Diehl JR, Harding PA 1997 Purification and characterization of novel heparin-binding growth factors in uterine secretory fluids: identification as heparin-regulated M_r 10,000 forms of connective tissue growth factor. J Biol Chem 272:20275–20282[Abstract/Free Full Text]
11. Ball DK, Surveyor GA, Diehl JR, Steffen CL, Uzumcu M, Mirando MA, Brigstock DR 1998 Characterization of 16- to 20- kilodalton (kDa) connective tissue growth factors (CTGFs) and demonstration of proteolytic activity for 38-kDa CTGF in pig uterine luminal flushings. Biol Reprod 59:828–835[Abstract/Free Full Text]
12. Kim H-S, Nagalla SR, Oh Y, Wilson E, Roberts Jr CT, Rosenfeld RG 1997 Identification of a family of low-affinity insulin-like growth factor binding proteins (IGFBPs): characterization of connective tissue growth factor as a member of the IGFBP superfamily. Proc Natl Acad Sci USA 94:12981–12986[Abstract/Free Full Text]
13. Bradham DM, Igarashi A, Potter RL, Grotendorst GR 1991 Connective tissue growth factor: a cysteine-rich mitogen secreted by human vascular endothelial cells is related to the SRC-induced immediate early gene product CEF-10. J Cell Biol 114:1285–1294[Abstract/Free Full Text]
14. Frazier K, Williams S, Kothapalli D, Klapper H, Grotendorst GR 1996 Stimulation of fibroblast cell growth, matrix production, and granulation tissue formation by connective tissue growth factor. J Invest Dermatol 107:404–411[CrossRef][Medline]
15. Kireeva ML, Latinki BV, Kolesnikova TV, Chen C-C, Yang GP, Abler AS, Lau LF 1997 Cyr61 and Fisp12 are both ECM-associated signaling molecules: activities, metabolism, and localization during development. Exp Cell Res 233:63–77[CrossRef][Medline]
16. Kireeva ML, Mo F-E, Yang GP, Lau LF 1996 Cyr61, a product of a growth factor-inducible immediate early gene, promotes cell proliferation, migration, and adhesion. Mol Cell Biol 16:1326–1334[Abstract/Free Full Text]
17. Kiefer MC, Masiarz FR, Bauer DM, Zapf J 1991 Identification and molecular cloning of two new 30-kDa insulin-like growth factor binding proteins isolated from adult human serum. J Biol Chem 266:9043–9049[Abstract/Free Full Text]
18. Oh Y, Nagalla SR, Yamanaka Y, Kim H-S, Wilson E, Rosenfled RG 1996 Synthesis and characterization of insulin-like growth factor-binding protein (IGFBP)-7. Recombinant human mac25 protein specificially binds IGF-I and II. J Biol Chem 271:30322–30325[Abstract/Free Full Text]
19. Burren CP, Wison E, Oh Y, Rosenfeld RG 1998 IGF binding demonstrated for NovH, a member of the CTGF family and the IGFBP superfamily. 80th Annual Meeting of The Endocrine Society, New Orleans, LA, 1998, P2–306 (Abstract)
20. Chevalier G, Yeger H, Martinerie C, Laurent M, Alami J, Schofield PN, Perbal B 1998 NovH: differential expression in developing kidney and Wilms tumors. Am J Pathol 152:1563–1575[Abstract]
21. Baxter RC, Binoux MA, Clemmons DR, Conover C, Drop SLS, Holly J, Mohan S, Oh Y, Rosenfeld RG 1998 Recommendations for nomenclature of the insulin-like growth factor binding protein superfamily. Endocrinology 139:4036[Free Full Text]
22. Collet C, Candy J 1998 How many insulin-like growth factor binding proteins? Mol Cell Endocrinol 139:1–6[CrossRef][Medline]
23. Voorberg J, Fontijn R, Calafat J, Janssen H, Van Mourik JA, Pannekoek H 1991 Assembly and routing of von Willebrand factor variants: the requirements for disulfide-linked dimerization reside within the carboxy-terminal 151 amino acids. J Cell Biol 113:195–205[Abstract/Free Full Text]
24. Pennica D, Swanson TA, Welsh JW, Roy MA, Lawrence DA, Lee J, Brush J, Taneyhill LA, Deuel B, Lew M, Watanabe C, Cohen RL, Melhem MF, Finley GG, Quirke P, Goddard AD, Hillan KJ, Gurney AL, Botstein D, Levine AJ 1998 WISP genes are members of the connective tissue growth factor family that are up-regulated in Wnt-1-transformed cells and aberrantly expressed in human colon tumors. Proc Natl Acad Sci USA 95:14717–14722[Abstract/Free Full Text]
25. Rich KA, George IV FW, Law JL, Martin WJ 1990 Cell-adhesive motif in region II of malarial circumsporozoite protein. Science 249:1574–1577[Abstract/Free Full Text]
26. Guo N-H, Krutzsch HC, Nègre E, Vogel T, Blake DA, Roberts DD 1992 Heparin- and sulfatide-binding peptides from the type 1 repeats of human thrombospondin promote melanoma cell adhesion. Proc Natl Acad Sci USA 89:3040–3044[Abstract/Free Full Text]
27. Holt GD, Pangburn MK, Ginsburg V 1990 Properdin binds to sulfatide [Gal(3-SO₄)ß1–1Cer] and has a sequence homology with other proteins that bind sulfated glycoconjugates. J Biol Chem 265:2852–2855[Abstract/Free Full Text]
28. McDonald NQ, Hendrickson WA 1993 A structural superfamily of growth factors containing a cystine knot motif. Cell 73:421–424[CrossRef][Medline]
29. Murzin AG, Chothia C 1992 Protein architecture: new superfamilies. Curr Opin Struct Biol 2:985–903
30. Almendral JM, Sommer D, MacDonald-Bravo H, Burckhardt J, Perera J, Bravo R 1988 Complexity of the early genetic response to growth factors in mouse fibroblasts. Mol Cell Biol 8:2140–2148[Abstract/Free Full Text]
31. Brunner A, Chinn J, Neubauer M, Purchio AF 1991 Identification of a gene family regulated by transforming growth factor-ß. DNA Cell Biol 10:293–300[Medline]
32. Harding PA, Surveyor GA, Brigstock DR 1998 Characterization of pig connective tissue growth factor (CTGF) cDNA, mRNA and protein from uterine tissue. DNA Seq 8:385–390[Medline]
33. Ying Z, Ling ML 1996 Isolation and characterization of xnov, a Xenopus laevis ortholog of the chicken nov gene. Gene 171:243–248[CrossRef][Medline]
34. Lasky JA, Ortiz LA, Tonthat B, Hoyle GW, Corti M, Athas G, Lungarella G, Brody A, Friedman M 1998 Connective tissue growth factor mRNA expression is upregulated in bleomycin-induced lung fibrosis. Am J Physiol 275:L365–L371
35. Lin J, Liliensiek B, Kanitz M, Schimanski U, Böhrer H, Waldherr R, Martin E, Kauffmann G, Ziegler R, Nawroth PP 1998 Molecular cloning of genes differentially regulated by TNF- in bovine aortic endothelial cells, fibroblasts and smooth muscle cells. Cardiovasc Res 38:802–803[Abstract/Free Full Text]
36. Grotendorst GR, Okochi H, Hayashi N 1996 A novel transforming growth factor ß response element controls the expression of the connective tissue growth factor gene. Cell Growth Differ 7:469–480[Abstract]
37. Xin LW, Martinerie C, Zumkeller W, Westphal M, Perbal B 1996 Differential expression of novH and CTGF in human glioma cell lines. Mol Pathol 49:M91–M97
38. Martinerie C, Viegas-Pequignot E, Guenard I, Dutrillaux B, Nguyen VC, Bernheim A, Perbal B 1992 Physical mapping of human loci homologous to the chicken nov proto-oncogene. Oncogene 7:2529–2534[Medline]
39. Steffen CL, Ball-Mirth DK, Harding PA, Bhattacharyya N, Pillai S, Brigstock DR 1998 Characterization of cell-associated and soluble forms of connective tissue growth factor (CTGF) produced by fibroblast cells in vitro. Growth Factors 15:199–213[Medline]
40. Yang D-H, Kim H-S, Wilson EM, Rosenfeld RG, Oh Y 1998 Identification of glycosylated 38-kDa connective tissue growth factor (IGFBP-related protein 2) and proteolytic fragments in human biological fluids, and up-regulation of IGFBP-rP2 expression by TGF-ß in Hs578T human breast cancer cells. J Clin Endocrinol Metab 83:2593–2596[Abstract/Free Full Text]
41. Igarashi A, Okochi H, Bradham DM, Grotendorst GR 1993 Regulation of connective tissue growth factor gene expression in human skin fibroblasts and during wound repair. Mol Biol Cell 4:637–645[Abstract]
42. Igarashi A, Nashiro K, Kikuchi K, Sato S, Ihn H, Grotendorst GR, Takehara K 1995 Significant correlation between connective tissue growth factor gene expression and skin sclerosis in tissue sections from patients with systemic sclerosis. J Invest Dermatol 105:280–284[CrossRef][Medline]
43. Shimo T, Nakanishi T, Kimura Y, Nishida T, Ishizeki K, Matsumura T, Takigawa T 1998 Inhibition of endogenous expression of connnetive tissue growth factor by its antisense oligonucleotide and antisense RNA supresses proliferation and migration of vascular endothelial cells. J Biochem 124:130–140[Abstract/Free Full Text]
44. Oemar BS, Werner A, Garnier J-M, Do D-D, Godoy N, Nauck M, März W, Rupp J, Pech M, Lüscher TF 1997 Human connective tissue growth factor is expressed in advanced atherosclerotic lesions. Circulation 95:831–839[Abstract/Free Full Text]
45. Hammes MS, Lieske JC, Pawar S, Spargo BH, Toback FG 1995 Calcium oxalate monohydrate crystals stimulate gene expression in renal epithelial cells. Kidney Int 48:501–509[Medline]
46. Pawar S, Kartha S, Toback FG 1995 Differential gene expression in migrating renal epithelial cells after wounding. J Cell Physiol 165:556–565[CrossRef][Medline]
47. Surveyor GA, Wilson AK, Brigstock DR 1998 Localization of connective tissue growth factor during the period of embryo implantation in the mouse. Biol Reprod 59:1207–1213[Abstract/Free Full Text]
48. Nakanishi T, Kimura Y, Tamura T, Ichikawa H, Yamaai Y, Sugimoto T, Takigawa M 1997 Cloning of a mRNA preferentially expressed in chondrocytes by differential display-PCR from a human chondrocytic cell line that is identical with connective tissue growth factor. Biochem Biophys Res Commun 234:206–210[CrossRef][Medline]
49. Kolesnikova TV, Lau LF 1998 Human CYR61-mediated enhancement of bFGF-induced DNA synthesis in human umbilical vein endothelial cells. Oncogene 16:747–754[CrossRef][Medline]
50. Burt DW 1992 Evolutionary grouping of the transforming growth factor-ß superfamily. Biochem Biophys Res Commun 184 590–595
51. Tamatani T, Kobayashi H, Tezuka K, Sakamoto S, Suzuki K, Nakanishi T, Takigawa M, Miyano T 1998 Establishment of the enzyme-linked immunosorbent assay for connective tissue growth factor (CTGF) and its detection in the sera of biliary atresia. Biochem Biophys Res Commun 251:748–752[CrossRef][Medline]
52. Kothapalli D, Hayashi N, Grotendorst GR 1998 Inhibition of TGF-ß-stimulated CTGF gene expression and anchorage-independent growth by cAMP identifies a CTGF-dependent restriction point in the cell cycle. FASEB J 12:1151–1161[Abstract/Free Full Text]
53. Grotendorst GR 1997 Connective tissue growth factor: a mediator of TGF-ß action on fibroblasts. Cytokine Growth Factor Rev 8:171–179[CrossRef][Medline]
54. Wenger C, Ellenrieder V, Alber B, Lacher U, Menke A, Hameister H, Wilda M, Iwamura T, Beger HG, Adler G, Gress TM 1999 Expression and differential regulation of connective tissue growth factor in pancreatic cancer cells. Oncogene 18:1073–1080[CrossRef][Medline]
55. Dammeier J, Beer H-D, Brauchle M, Werner S 1998 Dexamethasone is a novel potent inducer of connective tissue growth factor expression. Implications for glucocorticoid therapy. J Biol Chem 273:18185–18190[Abstract/Free Full Text]
56. Brigstock DR 1999 Purification and characterization of connective tissue growth factor (CTGF) using heparin-affinity chromatography. In: Aboul-Enein HY (ed) Analytical and Preparative Separation Methods of Biomacromolecules. Marcel Dekker Inc, New York, pp 397–414
57. Cardin AD, Weintraub HJR 1989 Molecular modeling of protein-glycosaminoglycan interactions. Arteriosclerosis 9:21–32[Abstract/Free Full Text]
58. McKeehan WL, Kan M 1994 Heparan sulfate fibroblast growth factor receptor complex: structure-function relationships. Mol Reprod Dev 39:69–82[CrossRef][Medline]
59. Yayon A, Klagsbrun M, Esko JD, Leder P, Ornitz DM 1991 Cell surface, heparin-like molecules are required for binding of basic fibroblast growth factor to its high affinity receptor. Cell 64:841–848[CrossRef][Medline]
60. Olwin BB, Rapraeger A 1992 Repression of myogenic differentiation by aFGF, bFGF and K-FGF is dependent on cellular heparan sulfate. J Cell Biol 118:631–639[Abstract/Free Full Text]
61. Rapraeger AC, Krufka A, Olwin BB 1991 Requirement of heparan sulfate for bFGF-mediated fibroblast growth and myoblast differentiation. Science 252:1705–1708[Abstract/Free Full Text]
62. Besner GE, Whelton D, Crissman-Combs MA, Steffen CL, Kim GY, Brigstock DR 1992 Interaction of heparin-binding EGF-like growth factor (HB-EGF) with the epidermal growth factor receptor: modulation by heparin, heparinase, or synthetic heparin-binding HB-EGF fragments. Growth Factors 7:289–296[Medline]
63. Aviezer D, Yayon A 1994 Heparin-dependent binding and autophosphorylation of epidermal growth factor (EGF) receptor by heparin-binding EGF-like growth factor but not by EGF. Proc Natl Acad Sci USA 91:12173–12177[Abstract/Free Full Text]
64. Higashiyama S, Abraham JA, Klagsbrun M 1993 Heparin-binding EGF-like growth factor stimulation of smooth muscle cell migration: dependence on interactions with cell surface heparan sulfate. J Cell Biol 122:933–940[Abstract/Free Full Text]
65. Cook PW, Damm D, Garrick BL, Wood KM, Karkaria CE, Higashiyama S, Klagsbrun M, Abraham JA 1995 Carboxyl-terminal truncation of leucine 76 converts heparin-binding EGF-like growth factor from a heparin-enhancible to a heparin-suppressible growth factor. J Cell Physiol 163:407–417[CrossRef][Medline]
66. Cook PW, Mattox PA, Keeble WW, Pittelkow MR, Plowman GD, Shoyab M, Adelman JP, Shipley GD 1991 A heparin-sulfate regulated human keratinocyte autocrine factor is similar or identical to amphiregulin. Mol Cell Biol 11:2547–2557[Abstract/Free Full Text]
67. Piepkorn M, Lo C, Plowman G 1994 Amphiregulin-dependent proliferation of cultured human keratinocytes: autocrine growth, the effects of exogenous recombinant cytokine, and apparent requirement for heparin-like glycosaminoglycans. J Cell Physiol 159:114–120[CrossRef][Medline]
68. Li S, Plowman GD, Buckley SD, Shipley GD 1992 Heparin inhibition of autonomous growth implicates amphiregulin as an autocrine growth factor for normal human mammary epithelial cells. J Cell Physiol 153:103–111[CrossRef][Medline]
69. Ron D, Bottaro DP, Finch PW, Morris D, Rubin JS, Aaronson SA 1993 Expression of biologically active recombinant keratinocyte growth factor: structure/function analysis of amino-terminal truncation mutants. J Biol Chem 268:2984–2988[Abstract/Free Full Text]
70. Gitay-Goren H, Soker S, Vlodavsky I, Neufeld G 1992 The binding of vascular endothelial growth factor to its receptors is dependent on cell surface-associated heparin-like molecules. J Biol Chem 267:6093–6098[Abstract/Free Full Text]
71. Yang GP, Lau LF 1991 Cyr61, product of a growth factor-inducible immediate early gene, is associated with the extracellular matrix and the cell surface. Cell Growth Differ 2:351–357[Abstract]
72. Shinozaki M, Kawara S, Hayashi N, Kakinuma T, Igarashi A, Takehara K 1997 Induction of subcutaneous tissue fibrosis in newborn mice by transforming growth factor ß: simultaneous application with basic fibroblast growth factor causes persistent fibrosis. Biochem Biophys Res Commun 237:292–296[CrossRef][Medline]
73. Scuri M, Grotendorst GR, Glassberg MK 1997 Connective tissue growth factor stimulates proliferation of human lung fibroblasts. Am J Respir Crit Care Med 155 A313 (Abstract)
74. Kothapalli D, Frazier KS, Welply A, Segarini PR, Grotendorst GR 1997 Transforming growth factor ß induces anchorage-independent growth of NRK fibroblasts via a connective tissue growth factor-dependent pathway. Cell Growth Differ 8:61–68[Abstract]
75. Igarashi A, Bradham DM, Okochi H, Grotendorst GR 1992 Connective tissue growth factor. J Dermatol 19:642–643[Medline]
76. Nishida T, Nakanishi T, Shimo T, Asano M, Hattori T, Tamatani T, Tezuka K, Takigawa M 1998 Demonstration of receptors specific for connective tissue growth factor on a human chondrocytic cell line (HCS-2/8). Biochem Biophys Res Commun 247:905–909[CrossRef][Medline]
77. Lau LF, Nathans D 1985 Identification of a set of genes expressed during the G0/G1 transition of cultured mouse cells. EMBO J 4:3145–3151[Medline]
78. O’Brien TP, Yang GP, Sanders L, Lau LF 1990 Expression of cyr61, a growth factor-inducible immediate-early gene. Mol Cell Biol 10:3569–3577[Abstract/Free Full Text]
79. Simmons DL, Levy DB, Yannoni Y, Erickson RL 1989 Identification of a phorbol ester-repressive v-src-inducible gene. Proc Natl Acad Sci USA 86:1178–1182[Abstract/Free Full Text]
80. Jay P, Bergé-Lefranc JL, Marsollier C, Méjean C, Taviaux S, Berta P 1997 The human growth factor-inducible immediate early gene, CYR61, maps to chromosome 1p. Oncogene 14:1753–1757[CrossRef][Medline]
81. Lau LF, Nathans D 1987 Expression of a set of growth-regulated immediate early genes in BALB/c 3T3 cells: coordinate regulation with c-fos or c-myc. Proc Natl Acad Sci USA 84:1182–1186[Abstract/Free Full Text]
82. Martinerie C, Viegas-Pequignot E, Nguyen VC, Perbal B 1997 Chromosomal mapping and expression of the human cyr61 gene in tumour cells from the nervous system. Mol Pathol 50:310–316[Abstract/Free Full Text]
83. O’Brien TP, Lau LF 1992 Expression of the growth factor-inducible immediate early gene cyr61 correlates with chondrogenesis during mouse embryonic development. Cell Growth Differ 3:645–654[Abstract]
84. Chung KC, Ahn YS 1998 Expression of immediate early gene cyr61 during the differentiation of immortalized embryonic hippocampal neuronal cells. Neurosci Lett 255:155–158[CrossRef][Medline]
85. Schütze N, Lechner A, Groll C, Siggelkow H, Hüfner M, Köhrle J, Jakob F 1998 The human analog of murine cysteine rich protein 61 is a 1,25-dihydroxyvitamin D₃ responsive immediate early gene in human fetal osteoblasts: regulation by cytokines, growth factors, and serum. Endocrinology 139:1761–1770[Abstract/Free Full Text]
86. Rivera-Gonzalez R, Peterson DN, Tkalcevic G, Thompson DD, Brown TA 1998 Estrogen-induced genes in the uterus of ovariectomized rats and their regulation by droloxifene and tamoxifen. J Steroid Biochem Mol Biol 64:13–24[CrossRef][Medline]
87. Babic AM, Kireeva ML, Kolesnikova TV, Lau LF 1998 CYR61, a product of a growth factor-inducible immediate early gene, promotes angiogenesis and tumor growth. Proc Natl Acad Sci USA 95:6355–6360[Abstract/Free Full Text]
88. Wong M, Kireeva ML, Kolesnikova TV, Lau LF 1997 Cyr61, product of a growth factor-inducible immediate-early gene, regulates chondrogenesis in mouse limb bud mesenchymal cells. Dev Biol 192:492–508[CrossRef][Medline]
89. Kireeva ML, Lam S C-T, Lau LF 1998 Adhesion of human umbilical vein endothelial cells to the immediate-early gene product Cyr61 in mediated through integrin _vß₃. J Biol Chem 273:3090–3096[Abstract/Free Full Text]
90. Clark EA, Brugge JS 1995 Integrins and signal transduction pathways: the road taken. Science 268:233–239[Abstract/Free Full Text]
91. Martinerie C, Perbal B 1991 Expression of a gene encoding a novel potential IGF binding protein in human tissues. C R Acad Sci III 313:345–351[Medline]
92. Snaith MR, Natarajan D, Taylor LB, Choi C-P, Martinerie C, Perbal B, Schofield PN, Boulter CA 1996 Genomic structure and chromosomal mapping of the mouse nov gene. Genomics 38:425–428[CrossRef][Medline]
93. Perbal B 1994 Contribution of MAV-1-induced nephroblastoma to the study of genes involved in human Wilms’ tumor development. Crit Rev Oncog 5:589–613[Medline]
94. Martinerie C, Chevalier G, Rauscher III FJ, Perbal B 1996 Regulation of nov by WT1: a potential role in nephrogenesis. Oncogene 12:1479–1492[Medline]
95. Scholz G, Martinerie C, Perbal B, Hanafusa H 1996 Transcriptional down regulation of the nov proto-oncogene in fibroblasts transformed by p60^v-src. Mol Cell Biol 16:481–486[Abstract/Free Full Text]
96. Hashimoto Y, Shindo-Okada N, Tani M, Takeuchi K, Toma H, Yokota J 1996 Identification of genes differentially expressed in association with metastatic potential of K-1735 murine melanoma by messenger RNA differential display. Cancer Res 56:5266–5271[Abstract/Free Full Text]
97. Hashimoto Y, Shindo-Okada N, Tani M, Nagamchi Y, Takeuchi K, Shiroishi T, Toma H, Yokota J 1998 Expression of the Elm1 gene, a novel gene of the CCN (connective tissue growth factor, cyr61/cef-10 and neuroblastoma overexpressed gene) family, suppresses in vivo tumor growth and metastasis of K-1735 murine melanoma cells. J Exp Med 3:289–296
98. Delmolino LM, Stearns NA, Castellot JJ 1997 Heparin induces a novel member of the CCN family which has characteristics of a growth arrest gene. Mol Biol Cell [Suppl] 8:A1665 (Abstract)
99. Zhang R, Averboukh L, Zhu W, Zhang H, Jo H, Dempsey PJ, Coffey RJ, Pardee AB, Liang P 1998 Identification of rCop-1, a new member of the CCN protein family, as a negative regulator for cell transformation. Mol Cell Biol 18:6131–6141[Abstract/Free Full Text]
100. Ebner R, Chopra A, Ruben SM 1998 Connective tissue growth factor-3. International patent WO 98/21236
101. Mason ED, Konrad KD, Webb CD, Marsh JL 1994 Dorsal midline fate in Drosophila embryos requires twisted gastrulation, a gene encoding a secreted protein related to human connective tissue growth factor. Genes Dev 8:1489–1501[Abstract/Free Full Text]
102. François V, Solloway M, O’Neill JW, Emery J, Bier E 1994 Dorsal-ventral patterning of the Drosophila embryo depends on a putative negative growth factor encoded by the short gastrulation gene. Genes Dev 8:2602–2616[Abstract/Free Full Text]
103. Mason ED, Williams S, Grotendorst GR, Marsh JL 1997 Combinatorial signaling by twisted gastrulation and decapentaplegic. Mech Dev 64:61–75[CrossRef][Medline]
104. Hastings GA, Adams MD 1998 Human CCN-like growth factor. US patent 5,780,263
105. Herschman HR 1991 Primary response genes induced by growth factors and tumor promoters. Annu Rev Biochem 60:381–319
106. Fornace Jr AJ, Jackman J, Hollander MC, Hoffmann-Liebermann B, Liebermann DA 1992 Genotoxic-stress-response genes and growth-arrest genes. gadd, MyD, and other genes induced by treatments eliciting growth arrest. Ann NY Acad Sci 663:139–153[Medline]
107. Fleming JV, Fontanier N, Harries DN, Rees WD 1997 The growth arrest genes gas5, gas6 and CHOP-10 (gadd153) are expressed in the mouse preimplantation embryo. Mol Reprod Dev 48:310–316[CrossRef][Medline]
108. Shugart EC, Levenson AS, Constance CM, Umek RM 1995 Differential expression of gas and gadd genes at distinct growth arrest points during adipocyte development. Cell Growth Differ 6:1541–1547[Abstract]
109. Manzow S, Brancolini C, Marks F, Richter KH 1996 Expression of growth arrest-specific (Gas) genes in murine keratinocytes: Gas2 is specifically regulated. Exp Cell Res 224:200–203[CrossRef][Medline]
110. Mullins DE, Rifkin DB 1991 Induction of proteases and protease inhibitors by growth factors. In: Sporn MB, Roberts AB (eds) Peptide Growth Factors and Their Receptors. Springer-Verlag, New York, vol 2:481–507
111. Glasser SR, Lampelo S, Munir MI, Julian J 1987 Expression of desmin, laminin and fibronectin during in situ differentiation (decidualization) of rat uterine stromal cells. Differentiation 35:132–142[CrossRef][Medline]
112. Adams JC, Watt FM 1993 Regulation of development and differentiation by the extracellular matrix. Development 117:1183–1198[Medline]
113. Ekblom P, Lehtonen E, Saxen L, Timpl R 1981 Shift in collagen type as an early response to induction of the metanephric mesenchyme. J Cell Biol 89:276–283[Abstract/Free Full Text]
114. Brigstock DR, Heap RB, Brown KD 1989 Polypeptide growth factors in uterine tissues and secretions. J Reprod Fertil 85:747–758[Abstract/Free Full Text]
115. McLachlan JA, Nelson KG, Takahashi T, Bossert NL, Newbold RR, Korach KS 1990 Estrogens and growth factors in the development, growth and function of the female reproductive tract. In: Schomberg DW (ed) Growth Factors in Reproduction. Springer Verlag, New York, pp 197–203
116. Brigstock DR 1991 Growth factors in the uterus: steroidal regulation and biological actions. Baillieres Clin Endocrinol Metab 5:791–808[Medline]
117. Murphy LJ, Murphy LC 1994 Steroid hormone induction of growth factor and oncogene expression in the uterus. In: Khan SA, Stancel GM (eds) Protooncogenes and Growth Factors in Steroid Hormone Induced Growth and Differentiation CRC Press, Boca Raton, FL, pp 31–45
118. Ignar-Trowbridge DM, Pimentel M, Teng CT, Korach KS, McLachlan JA 1995 Cross talk between peptide growth factor and estrogen receptor signaling systems. Environ Health Perspect 103 [Suppl 7]:35–38
119. Cullingford TE, Pollard JW 1994 Growth factors as mediators of sex steroid action in the uterus during its preparation for implantation. In: Khan SA, Stancel GM (eds) Protooncogenes and Growth Factors in Steroid Hormone Induced Growth and Differentiation. CRC Press, Ann Arbor, MI, pp 13–30
120. Pollard JW 1990 Regulation of polypeptide growth factor synthesis and growth factor-related gene expression in the rat and mouse uterus before and after implantation. J Reprod Fertil 88:721–731[Abstract/Free Full Text]
121. Roberts RM, Bazer FW 1988 The functions of uterine secretions. J Reprod Fertil 82:875–892[CrossRef][Medline]
122. Uzumcu M, Al-Homsi MF, Brigstock DR, Ayberk H, Coskun S, Jaroudi K, Hollanders JMG 1998 Immunohistochemical localization of connective tissue growth factor in human endometrium and decidua. Biol Reprod 58[Suppl 1]:A400 (Abstract)
123. Gupta A, Bazer FW, Jaeger LA 1996 Differential expression of ß transforming growth factors (TGFß1, TGFß2, and TGFß3) and their receptors (Type I and Type II) in peri-implantation porcine conceptuses. Biol Reprod 55:796–802[Abstract]
124. Gupta A, Ing NH, Bazer FW, Bustamante LS, Jaeger LA 1998 ß- Transforming growth factors (TGFß) at the porcine conceptus-maternal interface. I. Expression of TGFß1, TGFß2, and TGFß3 messenger ribonucleic acids. Biol Reprod 59:905–910[Abstract/Free Full Text]
125. Gupta A, Dekaney CM, Bazer FW, Madrigal MM, Jaeger LA 1998 ß-Transforming growth factors (TGFß) at the porcine conceptus-maternal interface. II. Uterine TGFß bioactivity and expression of immunoreactive TGFßs (TGFß1, TGFß2, and TGFß3) and their receptors (Type I and Type II). Biol Reprod 59:911–917[Abstract/Free Full Text]
126. Kennedy TG 1979 Prostaglandins and increased endometrial vascular permeabilty resulting from the application of an artificial stimulus to the uterus of the rat sensitized for the decidual cell reaction. Biol Reprod 20:560–566[Abstract]
127. Moulton BC, Koenig BB 1985 Uterine deoxyribonucleic acid synthesis during preimplantation in precursors of stromal cell differentiation during decidualization. Endocrinology 115:1302–1307[Abstract/Free Full Text]
128. Wewer UM, Faber M, Liotta LA, Albrechtsen R 1985 Immunochemical and ultrastructural assessment of the nature of the pericellular basement membrane of human decidual cells. Lab Invest 53:624–633[Medline]
129. Roberts AB, Sporn MB 1991 The transforming growth factor-ßs. In: Sporn MB, Roberts AB (eds) Peptide Growth Factors and Their Receptors. Springer Verlag, New York, vol 1:419–472
130. Klagsbrun M, D’Amore PA 1991 Regulators of angiogenesis. Annu Rev Physiol 53:217–239[CrossRef][Medline]
131. Wong HL, Wahl SM 1991 Inflammation and repair. In: Sporn MB, Roberts AB (eds) Peptide Growth Factors and Their Receptors. Springer-Verlag, New York, vol 2:509–548
132. Nathans D, Lau LF, Christy B, Hartzell S, Nakabeppu RL, Ryder K 1989 The genomic response to growth factors. Cold Spring Harbor Symp Quant Biol 53:893–900
133. Schilling JA, Joel W, Shurley HM 1959 Wound healing: a comparative study of the histochemical changes in granulation tissue contained in steel mesh and polyvinyl sponge cylinders. Surgery 46:702–710
134. Frank S, Madlenar M, Werner S 1996 Transforming growth factors ß1, ß2, and ß3 and their receptors are differentially regulated during normal and impaired wound healing. J Biol Chem 271:10188–10193[Abstract/Free Full Text]
135. Roberts AB, Sporn MB, Assoian RK, Smith JM, Roche NS, Wakefiled LM, Heine UI, Liotta LA, Falanga V, Kehrl JH, Fauci AS 1986 Transforming growth factor type ß: rapid induction of fibrosis and angiogenesis in vivo and stimulation of collagen formation in vitro. Proc Natl Acad Sci USA 83:4167–4171[Abstract/Free Full Text]
136. Franklin TJ 1997 Therapeutic approaches to organ fibrosis. Int J Biochem Cell Biol 29:79–89[CrossRef][Medline]
137. Border WA, Noble NA 1994 Transforming growth factor-ß in tissue fibrosis. N Engl J Med 331:1286–1292[Free Full Text]
138. Igarashi A, Nashiro K, Kikuchi K, Sato S, Ihn H, Fujimoto M, Grotendorst GR, Takehara K 1996 Connective tissue growth factor gene expression in tissue sections from localized scleroderma, keloid, and other fibrotic skin disorders. J Invest Dermatol 106:729–733[CrossRef][Medline]
139. Ito Y, Aten J, Bende RJ, Oemar BS, Rabelink TJ, Weening JJ, Goldschmeding R 1998 Expression of connective tissue growth factor in human renal fibrosis. Kidney Int 53:853–861[CrossRef][Medline]
140. Ito Y, Kleij L, Aten J, Oemar BS, Weening JJ, Rabelink TJ, Goldschmeding R 1997 CTGF: expression in the rat Thy 1.1 model and regulation in mesangial cells and podocytes. J Am Soc Nephrol 8:517A (Abstract)[Abstract]
141. Murphy M, Godson C, Cannon S, Kato S, Mackenzie HS, Martin F, Brady HR 1999 Suppression subtractive hybridization identifies high glucose levels as a stimulus for expression of connective tissue growth factor and other genes in human mesangial cells. J Biol Chem 274:5830–5834[Abstract/Free Full Text]
142. Wahab NA, Harper K, Mason RM 1996 Expression of extracellular matrix molecules in human mesangial cells in response to prolonged hyperglycaemia. Biochem J 316:985–992
143. Baecher-Allan CM, Barth RK 1993 PCR analysis of cytokine induction profiles associated with mouse strain variation in susceptibility to pulmonary fibrosis. Reg Immunol 5:207–217[Medline]
144. Phan SH, Kunkel SL 1992 Lung cytokine production in bleomycin-induced pulmonary fibrosis. Exp Lung Res 18:29–43[Medline]
145. Shi-Wen X, Abraham DJ, Pantelides P, Martin GR, du Bois RM, Black CM 1997 Connective tissue growth factor in scleroderma-associated lung. Thorax 52[Suppl 6]:S79 (Abstract)
146. Oemar BS, Lüscher TF 1997 Connective tissue growth factor. Friend or foe? Arterioscler Thromb Vasc Biol 17:1483–1489[Abstract/Free Full Text]
147. Dammeier J, Brauchle M, Falk W, Grotendorst GR, Werner S 1998 Connective tissue growth factor: a novel regulator of mucosal repair and fibrosis in inflammatory bowel disease? Int J Biochem Cell Biol 30:909–922[CrossRef][Medline]
148. Frazier KS, Grotendorst GR 1997 Expression of connective tissue growth factor mRNA in the fibrous stroma of mammary tumors. Int J Biochem Cell Biol 29:153–161[CrossRef][Medline]
149. Kubo M, Kikuchi K, Nashiro K, Kakinuma T, Hayashi N, Nanko H, Tamaki K 1998 Expression of fibrogenic cytokines in desmoplastic malignant melanoma. Br J Dermatol 139:192–197[CrossRef][Medline]
150. Genini M, Schwalbe P, Scholl FA, Schäfer BW 1996 Isolation of genes differentially expressed in human primary myoblasts and embryonal rhabdomyosarcoma. Int J Cancer 66:571–577[CrossRef][Medline]
151. Igarashi A, Hayashi N, Nashiro K, Takehara K 1998 Differential expression of connective tissue growth factor gene in cutaneous fibrohistiocytic and vascular tumors. J Cutan Pathol 25:143–148[Medline]
152. Pilarsky CP, Schmidt U, Eifsrich C, Stade J, Froschermaier SE, Haase M, Faller G, Kirchner TW, Wirth MP 1998 Expression of the extracellular matrix signaling molecule Cyr61 is downregulated in prostate cancer. Prostate 36:85–91[CrossRef][Medline]
153. Flaumenhaft R, Rifkin DB 1992 The extracellular regulation of growth factor action. Mol Biol Cell 3:1057–1065[Medline]
154. Bornstein P 1995 Diversity of function is inherent in matricellular proteins: an appraisal of thrombospondin1. J Cell Biol 130:503–506[Free Full Text]
155. Perbal B, Martinerie C, Sainson R, Werner M, He B, Roizman B 1999 The C-terminal domain of the regulatory protein NOVH is sufficient to promote interaction with fibulin 1C: a clue for a role of NOVH in cell-adhesion signaling. Proc Natl Acad Sci USA 96:869–874

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				N. Chen, S.-J. Leu, V. Todorovic, S. C.-T. Lam, and L. F. Lau Identification of a Novel Integrin {alpha}v{beta}3 Binding Site in CCN1 (CYR61) Critical for Pro-angiogenic Activities in Vascular Endothelial Cells J. Biol. Chem., October 15, 2004; 279(42): 44166 - 44176. [Abstract] [Full Text] [PDF]

				S.-J. Leu, N. Chen, C.-C. Chen, V. Todorovic, T. Bai, V. Juric, Y. Liu, G. Yan, S. C.-T. Lam, and L. F. Lau Targeted Mutagenesis of the Angiogenic Protein CCN1 (CYR61): SELECTIVE INACTIVATION OF INTEGRIN {alpha}6{beta}1-HEPARAN SULFATE PROTEOGLYCAN CORECEPTOR-MEDIATED CELLULAR FUNCTIONS J. Biol. Chem., October 15, 2004; 279(42): 44177 - 44187. [Abstract] [Full Text] [PDF]

				A. M. Ramirez, S. Takagawa, M. Sekosan, H. A. Jaffe, J. Varga, and J. Roman Smad3 Deficiency Ameliorates Experimental Obliterative Bronchiolitis in a Heterotopic Tracheal Transplantation Model Am. J. Pathol., October 1, 2004; 165(4): 1223 - 1232. [Abstract] [Full Text] [PDF]

				F. K. Askari, R. Dick, M. Mao, and G. J. Brewer Tetrathiomolybdate Therapy Protects Against Concanavalin A and Carbon Tetrachloride Hepatic Damage in Mice Exp Biol Med, September 1, 2004; 229(8): 857 - 863. [Abstract] [Full Text] [PDF]

				D. M. French, R. J. Kaul, A. L. D'Souza, C. W. Crowley, M. Bao, G. D. Frantz, E. H. Filvaroff, and L. Desnoyers WISP-1 Is an Osteoblastic Regulator Expressed During Skeletal Development and Fracture Repair Am. J. Pathol., September 1, 2004; 165(3): 855 - 867. [Abstract] [Full Text] [PDF]

				C. T. Fu, J. F. Bechberger, M. A. Ozog, B. Perbal, and C. C. Naus CCN3 (NOV) Interacts with Connexin43 in C6 Glioma Cells: POSSIBLE MECHANISM OF CONNEXIN-MEDIATED GROWTH SUPPRESSION J. Biol. Chem., August 27, 2004; 279(35): 36943 - 36950. [Abstract] [Full Text] [PDF]

				H. Yokoi, M. Mukoyama, T. Nagae, K. Mori, T. Suganami, K. Sawai, T. Yoshioka, M. Koshikawa, T. Nishida, M. Takigawa, et al. Reduction in Connective Tissue Growth Factor by Antisense Treatment Ameliorates Renal Tubulointerstitial Fibrosis J. Am. Soc. Nephrol., June 1, 2004; 15(6): 1430 - 1440. [Abstract] [Full Text] [PDF]

				S. Sakamoto, M. Yokoyama, X. Zhang, K. Prakash, K. Nagao, T. Hatanaka, R. H. Getzenberg, and Y. Kakehi Increased Expression of CYR61, an Extracellular Matrix Signaling Protein, in Human Benign Prostatic Hyperplasia and Its Regulation by Lysophosphatidic Acid Endocrinology, June 1, 2004; 145(6): 2929 - 2940. [Abstract] [Full Text] [PDF]

				Y. Absenger, H. Hess-Stumpp, B. Kreft, J. Kratzschmar, B. Haendler, N. Schutze, P.-A. Regidor, and E. Winterhager Cyr61, a deregulated gene in endometriosis Mol. Hum. Reprod., June 1, 2004; 10(6): 399 - 407. [Abstract] [Full Text] [PDF]

				E. M. Tam, C. J. Morrison, Y. I. Wu, M. S. Stack, and C. M. Overall Membrane protease proteomics: Isotope-coded affinity tag MS identification of undescribed MT1-matrix metalloproteinase substrates PNAS, May 4, 2004; 101(18): 6917 - 6922. [Abstract] [Full Text] [PDF]

				P. C. Trackman and A. Kantarci CONNECTIVE TISSUE METABOLISM AND GINGIVAL OVERGROWTH Critical Reviews in Oral Biology & Medicine, May 1, 2004; 15(3): 165 - 175. [Abstract] [Full Text] [PDF]

				A W Rachfal, M H Luquette, and D R Brigstock Expression of connective tissue growth factor (CCN2) in desmoplastic small round cell tumour J. Clin. Pathol., April 1, 2004; 57(4): 422 - 425. [Abstract] [Full Text] [PDF]

				D. Xie, D. Yin, H.-J. Wang, G.-T. Liu, R. Elashoff, K. Black, and H. P. Koeffler Levels of Expression of CYR61 and CTGF Are Prognostic for Tumor Progression and Survival of Individuals with Gliomas Clin. Cancer Res., March 15, 2004; 10(6): 2072 - 2081. [Abstract] [Full Text] [PDF]

				R. Gao and D. R. Brigstock Connective Tissue Growth Factor (CCN2) Induces Adhesion of Rat Activated Hepatic Stellate Cells by Binding of Its C-terminal Domain to Integrin {alpha}v{beta}3 and Heparan Sulfate Proteoglycan J. Biol. Chem., March 5, 2004; 279(10): 8848 - 8855. [Abstract] [Full Text] [PDF]

				C.-C. Chang, J.-Y. Shih, Y.-M. Jeng, J.-L. Su, B.-Z. Lin, S.-T. Chen, Y.-P. Chau, P.-C. Yang, and M.-L. Kuo Connective Tissue Growth Factor and Its Role in Lung Adenocarcinoma Invasion and Metastasis J Natl Cancer Inst, March 3, 2004; 96(5): 364 - 375. [Abstract] [Full Text] [PDF]

				S. Croci, L. Landuzzi, A. Astolfi, G. Nicoletti, A. Rosolen, F. Sartori, M. Y. Follo, N. Oliver, C. De Giovanni, P. Nanni, et al. Inhibition of Connective Tissue Growth Factor (CTGF/CCN2) Expression Decreases the Survival and Myogenic Differentiation of Human Rhabdomyosarcoma Cells Cancer Res., March 1, 2004; 64(5): 1730 - 1736. [Abstract] [Full Text] [PDF]

				M. Minato, S. Kubota, H. Kawaki, T. Nishida, A. Miyauchi, H. Hanagata, T. Nakanishi, T. Takano-Yamamoto, and M. Takigawa Module-Specific Antibodies against Human Connective Tissue Growth Factor: Utility for Structural and Functional Analysis of the Factor as Related to Chondrocytes J. Biochem., March 1, 2004; 135(3): 347 - 354. [Abstract] [Full Text] [PDF]

				H. R. Mason, A. C. Lake, J. E. Wubben, R. A. Nowak, and J. J. Castellot Jr The growth arrest-specific gene CCN5 is deficient in human leiomyomas and inhibits the proliferation and motility of cultured human uterine smooth muscle cells Mol. Hum. Reprod., March 1, 2004; 10(3): 181 - 187. [Abstract] [Full Text] [PDF]

				H. R. Mason, D. Grove-Strawser, B. S. Rubin, R. A. Nowak, and J. J. Castellot Jr. Estrogen Induces CCN5 Expression in the Rat Uterus in Vivo Endocrinology, February 1, 2004; 145(2): 976 - 982. [Abstract] [Full Text] [PDF]

				S. Sakamoto, M. Yokoyama, K. Prakash, J.-I. Tsuruha, S. Masamoto, R. H. Getzenberg, and Y. Kakehi Development of Quantitative Detection Assays for CYR61 as a New Marker for Benign Prostatic Hyperplasia J Biomol Screen, December 1, 2003; 8(6): 701 - 711. [Abstract] [PDF]

				P. Bonniaud, P. J. Margetts, M. Kolb, T. Haberberger, M. Kelly, J. Robertson, and J. Gauldie Adenoviral Gene Transfer of Connective Tissue Growth Factor in the Lung Induces Transient Fibrosis Am. J. Respir. Crit. Care Med., October 1, 2003; 168(7): 770 - 778. [Abstract] [Full Text] [PDF]

				G. Weston, A. C. Trajstman, C. E. Gargett, U. Manuelpillai, B. J. Vollenhoven, and P. A.W. Rogers Fibroids display an anti-angiogenic gene expression profile when compared with adjacent myometrium Mol. Hum. Reprod., September 1, 2003; 9(9): 541 - 549. [Abstract] [Full Text] [PDF]

				Y. Liang, C. Li, V. M. Guzman, A. J. Evinger III, C. E. Protzman, A. H.-P. Krauss, and D. F. Woodward Comparison of Prostaglandin F2{alpha}, Bimatoprost (Prostamide), and Butaprost (EP2 Agonist) on Cyr61 and Connective Tissue Growth Factor Gene Expression J. Biol. Chem., July 11, 2003; 278(29): 27267 - 27277. [Abstract] [Full Text] [PDF]

				J. M. Schober, L. F. Lau, T. P. Ugarova, and S. C.-T. Lam Identification of a Novel Integrin {alpha}M{beta}2 Binding Site in CCN1 (CYR61), a Matricellular Protein Expressed in Healing Wounds and Atherosclerotic Lesions J. Biol. Chem., July 3, 2003; 278(28): 25808 - 25815. [Abstract] [Full Text] [PDF]

				S. WERNER and R. GROSE Regulation of Wound Healing by Growth Factors and Cytokines Physiol Rev, July 1, 2003; 83(3): 835 - 870. [Abstract] [Full Text] [PDF]

				P. Finckenberg, K. Inkinen, J. Ahonen, S. Merasto, M. Louhelainen, H. Vapaatalo, D. Muller, D. Ganten, F. Luft, and E. Mervaala Angiotensin II Induces Connective Tissue Growth Factor Gene Expression via Calcineurin-Dependent Pathways Am. J. Pathol., July 1, 2003; 163(1): 355 - 366. [Abstract] [Full Text] [PDF]

				C. G. Lin, S.-J. Leu, N. Chen, C. M. Tebeau, S.-X. Lin, C.-Y. Yeung, and L. F. Lau CCN3 (NOV) Is a Novel Angiogenic Regulator of the CCN Protein Family J. Biol. Chem., June 20, 2003; 278(26): 24200 - 24208. [Abstract] [Full Text] [PDF]

				K. Sawai, K. Mori, M. Mukoyama, A. Sugawara, T. Suganami, M. Koshikawa, K. Yahata, H. Makino, T. Nagae, Y. Fujinaga, et al. Angiogenic Protein Cyr61 is Expressed by Podocytes in Anti-Thy-1 Glomerulonephritis J. Am. Soc. Nephrol., May 1, 2003; 14(5): 1154 - 1163. [Abstract] [Full Text] [PDF]

				K. H. Kim, Y. K. Min, J.-H. Baik, L. F. Lau, B. Chaqour, and K. C. Chung Expression of Angiogenic Factor Cyr61 during Neuronal Cell Death via the Activation of c-Jun N-terminal Kinase and Serum Response Factor J. Biol. Chem., April 11, 2003; 278(16): 13847 - 13854. [Abstract] [Full Text] [PDF]

				A. Leask, A. Holmes, C. M. Black, and D. J. Abraham Connective Tissue Growth Factor Gene Regulation. REQUIREMENTS FOR ITS INDUCTION BY TRANSFORMING GROWTH FACTOR-beta 2 IN FIBROBLASTS J. Biol. Chem., April 4, 2003; 278(15): 13008 - 13015. [Abstract] [Full Text] [PDF]

				B Perbal, D R Brigstock, and L F Lau Report on the second international workshop on the CCN family of genes Mol. Pathol., April 1, 2003; 56(2): 80 - 85. [Abstract] [Full Text] [PDF]

				D R Brigstock, R Goldschmeding, K-i Katsube, S C-T Lam, L F Lau, K Lyons, C Naus, B Perbal, B Riser, M Takigawa, et al. Proposal for a unified CCN nomenclature Mol. Pathol., April 1, 2003; 56(2): 127 - 128. [Abstract] [Full Text] [PDF]

				H. Thibout, C. Martinerie, C. Creminon, F. Godeau, P. Boudou, Y. Le Bouc, and M. Laurent Characterization of Human NOV in Biological Fluids: An Enzyme Immunoassay for the Quantification of Human NOV in Sera from Patients with Diseases of the Adrenal Gland and of the Nervous System J. Clin. Endocrinol. Metab., January 1, 2003; 88(1): 327 - 336. [Abstract] [Full Text] [PDF]

				A. C. Lake, A. Bialik, K. Walsh, and J. J. Castellot Jr CCN5 Is a Growth Arrest-Specific Gene That Regulates Smooth Muscle Cell Proliferation and Motility Am. J. Pathol., January 1, 2003; 162(1): 219 - 231. [Abstract] [Full Text] [PDF]

				F.-E Mo, A. G. Muntean, C.-C. Chen, D. B. Stolz, S. C. Watkins, and L. F. Lau CYR61 (CCN1) Is Essential for Placental Development and Vascular Integrity Mol. Cell. Biol., December 15, 2002; 22(24): 8709 - 8720. [Abstract] [Full Text] [PDF]

				Y.-P. Cheon, Q. Li, X. Xu, F. J. DeMayo, I. C. Bagchi, and M. K. Bagchi A Genomic Approach to Identify Novel Progesterone Receptor Regulated Pathways in the Uterus during Implantation Mol. Endocrinol., December 1, 2002; 16(12): 2853 - 2871. [Abstract] [Full Text] [PDF]

				S.-J. Leu, S. C.-T. Lam, and L. F. Lau Pro-angiogenic Activities of CYR61 (CCN1) Mediated through Integrins alpha vbeta 3 and alpha 6beta 1 in Human Umbilical Vein Endothelial Cells J. Biol. Chem., November 22, 2002; 277(48): 46248 - 46255. [Abstract] [Full Text] [PDF]

				J. K. G. Crean, D. Finlay, M. Murphy, C. Moss, C. Godson, F. Martin, and H. R. Brady The Role of p42/44 MAPK and Protein Kinase B in Connective Tissue Growth Factor Induced Extracellular Matrix Protein Production, Cell Migration, and Actin Cytoskeletal Rearrangement in Human Mesangial Cells J. Biol. Chem., November 8, 2002; 277(46): 44187 - 44194. [Abstract] [Full Text] [PDF]

				J. Lafont, M. Laurent, H. Thibout, F. Lallemand, Y. Le Bouc, A. Atfi, and C. Martinerie The Expression of novH in Adrenocortical Cells Is Down-regulated by TGFbeta 1 through c-Jun in a Smad-independent Manner J. Biol. Chem., October 18, 2002; 277(43): 41220 - 41229. [Abstract] [Full Text] [PDF]

				B. Chaqour, C. Whitbeck, J.-S. Han, E. Macarak, P. Horan, P. Chichester, and R. Levin Cyr61 and CTGF are molecular markers of bladder wall remodeling after outlet obstruction Am J Physiol Endocrinol Metab, October 1, 2002; 283(4): E765 - E774. [Abstract] [Full Text] [PDF]

				C. R. Harlow, L. Davidson, K. H. Burns, C. Yan, M. M. Matzuk, and S. G. Hillier FSH and TGF-{beta} Superfamily Members Regulate Granulosa Cell Connective Tissue Growth Factor Gene Expression in Vitro and in Vivo Endocrinology, September 1, 2002; 143(9): 3316 - 3325. [Abstract] [Full Text] [PDF]

				K. J. Way, K. Isshiki, K. Suzuma, T. Yokota, D. Zvagelsky, F. J. Schoen, G. E. Sandusky, P. A. Pechous, C. J. Vlahos, H. Wakasaki, et al. Expression of Connective Tissue Growth Factor Is Increased in Injured Myocardium Associated With Protein Kinase C {beta}2 Activation and Diabetes Diabetes, September 1, 2002; 51(9): 2709 - 2718. [Abstract] [Full Text] [PDF]

				K. Sakamoto, S. Yamaguchi, R. Ando, A. Miyawaki, Y. Kabasawa, M. Takagi, C. L. Li, B. Perbal, and K.-i. Katsube The Nephroblastoma Overexpressed Gene (NOV/ccn3) Protein Associates with Notch1 Extracellular Domain and Inhibits Myoblast Differentiation via Notch Signaling Pathway J. Biol. Chem., August 9, 2002; 277(33): 29399 - 29405. [Abstract] [Full Text] [PDF]

				C L Li, V Martinez, B He, A Lombet, and B Perbal A role for CCN3 (NOV) in calcium signalling Mol. Pathol., August 1, 2002; 55(4): 250 - 261. [Abstract] [Full Text] [PDF]

				E E-D A Moussad, M A E Rageh, A K Wilson, R D Geisert, and D R Brigstock Temporal and spatial expression of connective tissue growth factor (CCN2; CTGF) and transforming growth factor {beta} type 1 (TGF-{beta}1) at the utero-placental interface during early pregnancy in the pig Mol. Pathol., June 1, 2002; 55(3): 186 - 192. [Abstract] [Full Text] [PDF]

				J. M. Schober, N. Chen, T. M. Grzeszkiewicz, I. Jovanovic, E. E. Emeson, T. P. Ugarova, R. D. Ye, L. F. Lau, and S. C.-T. Lam Identification of integrin alpha Mbeta 2 as an adhesion receptor on peripheral blood monocytes for Cyr61 (CCN1) and connective tissue growth factor (CCN2): immediate-early gene products expressed in atherosclerotic lesions Blood, May 29, 2002; 99(12): 4457 - 4465. [Abstract] [Full Text] [PDF]

				S. Kondo, S. Kubota, T. Shimo, T. Nishida, G. Yosimichi, T. Eguchi, T. Sugahara, and M. Takigawa Connective tissue growth factor increased by hypoxia may initiate angiogenesis in collaboration with matrix metalloproteinases Carcinogenesis, May 1, 2002; 23(5): 769 - 776. [Abstract] [Full Text] [PDF]

				T. M. Grzeszkiewicz, V. Lindner, N. Chen, S. C.-T. Lam, and L. F. Lau The Angiogenic Factor Cysteine-Rich 61 (CYR61, CCN1) Supports Vascular Smooth Muscle Cell Adhesion and Stimulates Chemotaxis through Integrin {alpha}6{beta}1 and Cell Surface Heparan Sulfate Proteoglycans Endocrinology, April 1, 2002; 143(4): 1441 - 1450. [Abstract] [Full Text] [PDF]

				M. C. Manara, B. Perbal, S. Benini, R. Strammiello, V. Cerisano, S. Perdichizzi, M. Serra, A. Astolfi, F. Bertoni, J. Alami, et al. The Expression of ccn3(nov) Gene in Musculoskeletal Tumors Am. J. Pathol., March 1, 2002; 160(3): 849 - 859. [Abstract] [Full Text] [PDF]

				J. Liu, V.-M. Kosma, T. Vanttinen, C. Hyden-Granskog, and R. Voutilainen Gonadotrophins inhibit the expression of insulin-like growth factor binding protein-related protein-2 mRNA in cultured human granulosa-luteal cells Mol. Hum. Reprod., February 1, 2002; 8(2): 136 - 141. [Abstract] [Full Text] [PDF]

				F. Su, M. Overholtzer, D. Besser, and A. J. Levine WISP-1 attenuates p53-mediated apoptosis in response to DNA damage through activation of the Akt kinase Genes & Dev., January 1, 2002; 16(1): 46 - 57. [Abstract] [Full Text] [PDF]

				C.-C. Chen, F.-E Mo, and L. F. Lau The Angiogenic Factor Cyr61 Activates a Genetic Program for Wound Healing in Human Skin Fibroblasts J. Biol. Chem., December 7, 2001; 276(50): 47329 - 47337. [Abstract] [Full Text] [PDF]

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