The tetraspanin CD9 influences the adhesion, spreading, and pericellular fibronectin matrix assembly of Chinese hamster ovary cells on human plasma fibronectin
The tetraspanin CD9 influences the adhesion, spreading, and pericellular fibronectin matrix assembly of Chinese hamster ovary cells on human plasma fibronectin
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DOI：
10.1006/excr.1999.4596
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发表时间：
1999-09-15
影响因子：
3.7
通讯作者：
Jennings, LK
中科院分区：
文献类型：
--
作者：
Cook, GA;Wilkinson, DA;Jennings, LK
使用转染的中国仓鼠卵巢（CHO）细胞系统研究了CD 9在细胞粘附和粘附蛋白上的铺展中的作用。CD 9细胞表面表达导致在纤连蛋白（Fn）上的粘附减少和铺展增加。而模拟转染（模拟CHO）和幼稚CHO细胞呈现典型的成纤维细胞纺锤形形态，CDS转染（CDS-CHO）细胞是多边形的许多丝状突起，并表现出两倍大的表面积。PB 1 mAb阻断α（5）β（1）可抑制CDS-CHO细胞（而非模拟CHO细胞）的扩散形态，表明α（5）β（1）和CD 9的共表达影响了细胞对Fn的活性。CD 9的第二细胞外环涉及粘附的调节，因为Fn上的CD 9-CHO细胞粘附减少通过抗CD 9抗体连接至第二细胞外环或与表达缺乏第二细胞外环结构域的CD 9突变体的细胞逆转。使用细胞粘附试验和ELISA，我们证明了CD 9与Fn的HEP 2/IIICS区域结合。最后，CD 9表达导致富含Fn的细胞周基质组装减少两倍。我们的观察结果表明，CD 9显着影响CHO细胞与Fn的相互作用，并表明CD 9在调节细胞-细胞外基质相互作用中具有重要作用。(C)北京：科学出版社.
The role of CD9 in cell adhesion and spreading on adhesive proteins was investigated using a transfected Chinese hamster ovary (CHO) cell system. CD9 cell surface expression resulted in reduced adhesion and increased spreading on fibronectin (Fn). Whereas mock-transfeeted (mock CHO) and naive CHO cells assumed a typical fibroblast spindle shape morphology, CDS-transfected (CDS-CHO) cells were polygonal with many filipodial projections and exhibited a twofold greater surface area. The spread morphology of CDS-CHO cells, but not mock CHO cells, was inhibited by PB1 mAb blockade of alpha(5)beta(1), suggesting that the coexpression of alpha(5)beta(1) and CD9 influenced cell activity on Fn. The second extracellular loop of CD9 was implicated in regulation of adhesion since reduced CD9-CHO cell adhesion on Fn was reversed by either anti-CD9 antibody Ligation to the second extracellular loop or with cells expressing a CD9 mutant lacking the second extracellular loop domain. Using cell adhesion assays and ELISA, we demonstrated CD9 binding to the HEP2/IIICS region of Fn. Finally, CD9 expression resulted in a twofold reduction in Fn-rich pericellular matrix assembly. Our observations show that CD9 dramatically influences CHO cell interactions with Fn and suggest that CD9 has an important role in modulating cell-extracellular matrix interactions. (C) 1999 Academic Press.