The interplay between Eps8 and IRSp53 contributes to Src-mediated transformation
The interplay between Eps8 and IRSp53 contributes to Src-mediated transformation
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DOI：
10.1038/onc.2010.144
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发表时间：
2010-07-08
期刊：
影响因子：
8
通讯作者：
Leu, T-H
中科院分区：
文献类型：
--
作者：
Liu, P-S;Jong, T-H;Leu, T-H
作为一种癌蛋白，Eps 8参与v-Src诱导的细胞转化。为了描述潜在的机制，我们进行了酵母双杂交筛选，并确定IRSp 53 S，细胞动员的关键蛋白质，作为Eps 8结合伴侣之一，从人脑cDNA文库。该协会是介导的多个脯氨酸丰富的区域Eps 8和C-末端SH 3-WWB含有结构域的IRSp 53 S。在这项研究中，我们观察到Eps 8调节v-Src转化细胞（IV 5）中IRSp 53的表达，提出了Eps 8/IRSp 53相互作用是否在癌发生中至关重要的问题。为了解决这个问题，我们产生了表达IV 5的irsp 53 siRNA细胞。IRSp 53的减毒可降低培养皿中IV 5的细胞增殖和小鼠肿瘤形成，这可以部分地被异位表达的人IRSp 53 S拯救。此外，IRSp 53敲低损害了磷脂酰肌醇3-激酶（如Pi-Ser 473 AKT所反映的）和Stat 3（如Pi-Tyr 705 Stat 3所反映的）的活性，并降低了细胞周期蛋白D1的表达，最终阻碍了G（1）期细胞周期的进展。异位表达的人IRSp 53 S，但不是其Eps 8结合缺陷突变体（即Delta 363和PPPDA），挽救了这些缺陷，并部分恢复细胞增殖。值得注意的是，通过激活Src，EGF增加了HeLa细胞中Eps 8/IRSp 53复合物的形成和Stat 3的激活。通过这些结果，我们首次表明IRSp 53通过与Eps 8的相互作用，不仅影响细胞迁移，而且还决定癌细胞的细胞生长。Oncogene（2010）29，3977-3989; doi：10.1038/onc.2010.144; 2010年4月26日在线发表
As an oncoprotein, Eps8 participates in v-Src-induced cellular transformation. To delineate the underlying mechanism, we conducted a yeast two-hybrid screening and identified IRSp53S, a protein critical in cell mobilization, as one of the Eps8-binding partners from a human brain cDNA library. The association was mediated by the multiple proline-rich regions of Eps8 and the C-terminal SH3-WWB containing domains of IRSp53S. In this study, we observed that Eps8 modulated the expression of IRSp53 in v-Src-transformed cells (IV5), raising the question of whether Eps8/IRSp53 interaction was crucial in carcinogenesis. To address this issue, we generated IV5-expressing irsp53 siRNA cells. Attenuation of IRSp53 reduced cell proliferation of IV5 in culture dish and tumor formation in mice, which could be partly rescued by ectopically expressed human IRSp53S. In addition, IRSp53 knockdown impaired activity of phosphatidylinositol 3-kinase (as reflected by Pi-Ser473 AKT) and Stat3 (as reflected by Pi-Tyr705 Stat3), and reduced cyclin D1 expression that culminated to impede G(1)-phase cell-cycle progression. Ectopically expressed human IRSp53S, but not its Eps8-binding defective mutants (that is, Delta 363 and PPPDA), rescued these defects and partly restored cell proliferation. Remarkably, through activation of Src, EGF increased the formation of Eps8/IRSp53 complex and Stat3 activation in HeLa cells. With these results, we show for the first time that IRSp53, through its interaction with Eps8, not only affects cell migration but also dictates cellular growth in cancer cells. Oncogene (2010) 29, 3977-3989; doi:10.1038/onc.2010.144; published online 26 April 2010