The catalytic activity of the Src family kinases is required to disrupt cadherin-dependent cell-cell contacts
The catalytic activity of the Src family kinases is required to disrupt cadherin-dependent cell-cell contacts
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DOI：
10.1091/mbc.11.1.51
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发表时间：
2000-01-01
影响因子：
3.3
通讯作者：
Brunton, VG
中科院分区：
文献类型：
--
作者：
Owens, DW;McLean, GW;Brunton, VG
尽管上皮细胞接触在决定细胞行为中的重要性，但我们仍然缺乏对细胞间接触的组装和拆卸的详细了解。在这里，我们研究了Src家族激酶在体外上皮细胞接触中的催化活性的作用。与E-和P-钙粘蛋白一样，正常和肿瘤来源的人角质形成细胞的Ca 2+处理导致c-Yes（和c-Src和Fyn）以及它们的假定底物p120（CTN）被招募到细胞-细胞接触中。一种对Src家族激酶具有选择性的酪氨酸激酶抑制剂，PD 162531和一种干扰内源性Src激酶催化功能的显性抑制性c-Src蛋白诱导细胞-细胞接触和E-钙粘蛋白再分布，即使在通常不支持稳定细胞-细胞粘附的低Ca 2+条件下。延时显微镜表明，Src激酶抑制诱导稳定的瞬时形成的细胞间接触在低钙。此外，E-和P-钙粘蛋白特异性抗体的组合抑制细胞-细胞接触，表明钙粘蛋白参与。作为接触稳定的结果，正常细胞不能从高密度形成的上皮层中解离并在体外修复伤口，尽管单个细胞仍然能动。因此，钙粘蛋白依赖性接触可以通过高Ca 2+和通过在体外低（0.03 mM）Ca 2+中抑制Src活性来稳定。
Despite the importance of epithelial cell contacts in determining cell behavior, we still lack a detailed understanding of the assembly and disassembly of intercellular contacts. Here we examined the role of the catalytic activity of the Src family kinases at epithelial cell contacts in vitro. Like E- and P-cadherin, Ca2+ treatment of normal and tumor-derived human keratinocytes resulted in c-Yes (and c-Src and Fyn), as well as their putative substrate p120(CTN), being recruited to cell-cell contacts. A tyrosine kinase inhibitor with selectivity against the Src family kinases, PD162531, and a dominant-inhibitory c-Src protein that interferes with the catalytic function of the endogenous Src kinases induced cell- cell contact and E-cadherin redistribution, even in low Ca2+ which does not normally support stable cell- cell adhesion. Time-lapse microscopy demonstrated that Src kinase inhibition induced stabilization of transiently formed intercellular contacts in low Ca2+. Furthermore, a combination of E- and P-cadherin-specific antibodies suppressed cell-cell contact, indicating cadherin involvement. As a consequence of contact stabilization, normal cells were unable to dissociate from an epithelial sheet formed at high density and repair a wound in vitro, although individual cells were still motile. Thus, cadherin-dependent contacts can be stabilized both by high Ca2+ and by inhibiting Src activity in low (0.03 mM) Ca2+ in vitro.