Grant D S, Kinsella J L, Fridman R, Auerbach R, Piasecki B A, Yamada Y, Zain M, Kleinman H K
Laboratory of Developmental Biology, National Institute of Dental Research, National Institutes of Health, Bethesda, Maryland 20892.
J Cell Physiol. 1992 Dec;153(3):614-25. doi: 10.1002/jcp.1041530324.
Endothelial cells are known to bind to laminin, and two peptides derived from the laminin A (CTFALRGDNP) and B1 (CDPGYIGSR) chains block the capillary-like tube formation on a laminin-rich basement membrane matrix, Matrigel. In the present study, we have used various in vitro and in vivo assays to investigate the angiogenic-biologic effects of a third active site in the laminin A chain, CSRARKQAASIKVAVSADR (designated PA22-2) on endothelial cells. The SIKVAV-containing peptide was as active as the YIGSR-containing peptide for endothelial cell attachment but was less active than either the RGD-containing peptide or intact laminin. Endothelial cells seeded on this peptide appeared fibroblastic with many extended processes, unlike the normal cobblestone morphology observed on tissue culture plastic. In addition, in contrast to normal tube formation on Matrigel, short irregular structures formed, some of which penetrated the matrix and sprouting was more apparent. Analysis of endothelial cell conditioned media of cells cultured in the presence of this peptide indicated degradation of the Matrigel and zymograms demonstrated active collagenase IV (gelatinase) at 68 and 62 Kd. A murine in vivo angiogenesis assay and the chick yolk sac/chorioallantoic membrane assays with the peptide demonstrated increased endothelial cell mobilization, capillary branching, and vessel formation. These data suggest that the -SIKVAV-site may play an important role in initiating branching and formation of new capillaries from the parent vessels, a behavior that is observed in vivo in response to tumor growth or in the normal vascular response to injury.
已知内皮细胞可与层粘连蛋白结合，并且源自层粘连蛋白A链（CTFALRGDNP）和B1链（CDPGYIGSR）的两种肽可阻断富含层粘连蛋白的基底膜基质Matrigel上的毛细血管样管形成。在本研究中，我们使用了各种体外和体内试验来研究层粘连蛋白A链中第三个活性位点CSRARKQAASIKVAVSADR（命名为PA22-2）对内皮细胞的血管生成生物学效应。含SIKVAV的肽在内皮细胞附着方面与含YIGSR的肽活性相当，但活性低于含RGD的肽或完整的层粘连蛋白。接种在该肽上的内皮细胞呈现出成纤维细胞样，有许多延伸的突起，这与在组织培养塑料上观察到的正常鹅卵石形态不同。此外，与Matrigel上的正常管形成相反，形成了短的不规则结构，其中一些穿透了基质，并且芽生更为明显。对在该肽存在下培养的细胞的内皮细胞条件培养基的分析表明Matrigel降解，酶谱显示在68和62 Kd处有活性的胶原酶IV（明胶酶）。用该肽进行的小鼠体内血管生成试验和鸡胚卵黄囊/绒毛尿囊膜试验表明内皮细胞动员、毛细血管分支和血管形成增加。这些数据表明，-SIKVAV-位点可能在启动从母血管形成新毛细血管的分支和形成中起重要作用，这种行为在体内可观察到，是对肿瘤生长的反应或正常血管对损伤的反应。
