摘要
目的探讨Y-27632对胃癌SGC-7901细胞侵袭和迁移的影响,及其作用机制是否是通过对SRF的调控来实现的。方法将细胞分为3组:1)对照组;2)Y-27632通路阻断剂组;3)siRNA-SRF-1107干扰组。各组分别转染、加药,孵育48 h。用CCK-8法检测细胞增殖,Transwell小室和划痕实验检测细胞侵袭能力和迁移能力。Western blot法检测SRF、ROCK1、E-cadherin、β-catenin、F-actin、MRTF-A及Cyclin D1的表达。结果 Y-27632能显著抑制胃癌SGC-7901细胞的侵袭、迁移能力(P<0.05),而对其增殖没有影响。Y-27632能显著下调SGC-7901细胞的ROCK1、SRF、F-actin和MRTF-A的表达,上调E-cadherin的表达(P<0.05)。结论 Y-27632通过上调E-cadherin表达,同时阻断ROCK信号抑制SRF辅因子MRTF-A的表达,最终下调SRF转录水平,从而抑制胃癌SGC-7901细胞侵袭、转移和迁移能力。
Objective To study the effect of Y-27632 on invasion and motility of SGC-7901 gastric carcinoma cells, and to find whether Y-27632 excerts the effect by attenuating SRF expression. Methods SGC-7901 gastric carcinoma cells were divided into 3 groups ： 1 ） blank control group ; 2） Y-27632 group ; 3 ） siRNA-SRF- 1107 group. Transfected siRNA-SRF or incubated by Y-27632 48 h. The effect of Y-27632 on proliferation suppressions of SGC-7901 gastric carcinoma cells was detected by CCK-8 assay. Cell invasion was examined by Transwell and wound healing test. The expression of SRF, ROCK1, E-cadherin, 13-catenin, F-actin, MRTF-A and Cyclin D1 were detected by Western blot. Results Y-27632 inhibited invasion （P 〈 0.05） but had no effect on proliferation of SGC-7901 gastric carcinoma cells. Y-27632 reduced ROCK1, MRTF-A, F-aetin, SRF protein expressions by 37.0% , 44.3% , 62.7% and 62. 7% respectively, and E-cadherin protein expression was up-regulated by 2.64 folds （P 〈 0.05 ）. Conclusions The inhibition of ROCK and up-regulation of E-eadherin by Y-27632 can inhibit the invasion and migration of SGC-7901 gastric carcinoma cells that is explained at least, in part, by attenuating SRF expression.
出处
《基础医学与临床》
CSCD
2015年第10期1369-1374,共6页
Basic and Clinical Medicine
基金
秦皇岛市科技支撑项目(201501B045)
