The angiogenic factors Cyr61 and connective tissue growth factor induce adhesive signaling in primary human skin fibroblasts
The angiogenic factors Cyr61 and connective tissue growth factor induce adhesive signaling in primary human skin fibroblasts
复制标题
DOI：
10.1074/jbc.m008087200
复制
发表时间：
2001-03-30
影响因子：
4.8
通讯作者：
Lau, LF
中科院分区：
文献类型：
--
作者：
Chen, CC;Chen, NY;Lau, LF
血管生成诱导剂富含半胱氨酸的血管生成蛋白61 （Cyr61）和结缔组织生长因子（CTGF）是结构相关的、细胞外基质相关的肝素结合蛋白。两者都能刺激内皮细胞和成纤维细胞的趋化和增殖，并在体内诱导新生血管。Cyr61和CTGF是由可诱导的直接早期基因编码的，在培养成纤维细胞和真皮成纤维细胞的皮肤伤口愈合过程中，生长因子刺激合成Cyr61和CTGF。最近，我们发现，原代人成纤维细胞与固定Cyr61的粘附是通过整合素cr、P和细胞表面硫酸肝素蛋白聚糖（HSPGs）介导的（Chen， N,, Chen， C， C, and Lau， L，F, (2000) J，生物化学，275,24953 -24961）。首次证明了HSPGs在整合素介导的细胞附着中是绝对需要的。我们在本研究中表明，CTGF也通过相同的机制介导成纤维细胞粘附，并证明成纤维细胞粘附于固定化Cyr61或CTGF诱导了与其生物活性相一致的不同的粘附信号反应。与成纤维细胞粘附纤维连接蛋白、层粘连蛋白或I型胶原相比，细胞粘附Cyr61或CTGF诱导1)丝状足和板足形成更广泛和更长时间，同时在假足前缘形成含整合素α (6) β(1)的局点复合物；2)细胞内信号分子的激活，包括黏附激酶、paxillin和Rac，具有类似的快速动力学；3) p42/p44 MAPKs持续激活至少9小时；4)延长的基因表达变化，包括MMP-1（胶原酶-1）和MMP-3（基质溶素-1）mrna和蛋白的上调，持续至少24小时。总之，这些结果表明Cyr61和CTGF是真正的具有特定信号能力的粘附底物。通过整合素α (6) β(1)和hspg介导的附着过程中的信号传导，为它们在成纤维细胞中的活性提供了分子基础，并表明这些蛋白可能在血管生成和伤口愈合过程中通过激活金属蛋白酶参与基质重塑。
The angiogenic inducers cysteine-rich angiogenic protein 61 (Cyr61) and connective tissue growth factor (CTGF) are structurally related, extracellular matrix-associated heparin-binding proteins. Both can stimulate chemotaxis and promote proliferation in endothelial cells and fibroblasts in culture and induce neovascularization in vivo. Encoded by inducible immediate early genes, Cyr61 and CTGF are synthesized upon growth factor stimulation in cultured fibroblasts and during cutaneous wound healing in dermal fibroblasts, Recently, we have shown that adhesion of primary human fibroblasts to immobilized Cyr61 is mediated through integrin cr,P, and cell surface heparan sulfate proteoglycans (HSPGs) (Chen, N,, Chen, C,-C,, and Lau, L,F, (2000) J, Biol, Chem, 275, 24953-24961), providing the first demonstration of an absolute requirement for HSPGs in integrin-mediated cell attachment. We show in this study that CTGF also mediates fibroblast adhesion through the same mechanism and demonstrate that fibroblasts adhesion to immobilized Cyr61 or CTGF induces distinct adhesive signaling responses consistent with their biological activities. Compared with fibroblast adhesion to fibronectin, laminin, or type I collagen, cell adhesion to Cyr61 or CTGF induces 1) more extensive and prolonged formation of filopodia and lamellipodia, concomitant with formation of integrin alpha (6)beta (1)-containing focal complexes localized at leading edges of pseudopods; 2) activation of intracellular signaling molecules including focal adhesion kinase, paxillin, and Rac with similar rapid kinetics; 3) sustained activation of p42/p44 MAPKs lasting for at least 9 h; and 4) prolonged gene expression changes including up-regulation of MMP-1 (collagenase-l) and MMP-3 (stromelysin-l) mRNAs and proteins sustained for at least 24 h, Together, these results establish Cyr61 and CTGF as bona fide adhesive substrates with specific signaling capabilities, provide a molecular basis for their activities in fibroblasts through integrin alpha (6)beta (1) and HSPG-mediated signaling during attachment and indicate that these proteins may function in matrix remodeling through the activation of metalloproteinases during angiogenesis and wound healing.