摘要
目的观察用RNA干扰技术沉默人胃癌细胞株MGC-803中钙结合蛋白S100A4的基因,对基质金属蛋白酶2(matrix metalloproteinase 2,MMP-2)和E-钙黏素(E-cadherin)表达变化,探讨S100A4对胃癌细胞侵袭能力的影响及作用机制。方法采用RNA干扰技术将针对S100A4和非特异性阴性对照序列的si RNA转染至人胃癌MGC-803细胞,利用荧光实时定量RT-PCR和蛋白免疫印迹法(Western Blot)检测S100A4基因和蛋白的表达,筛选S100A4基因沉默组和阴性对照组细胞。采用Western Blot方法检测MMP-2和E-cadherin蛋白表达。利用Transwell Matrigel方法检测各实验组中MGC-803细胞体外侵袭能力。结果成功建立S100A4基因沉默的人胃癌MGC-803细胞。S100A4基因沉默的胃癌MGC-803细胞中MMP-2蛋白表达降低,而E-cadherin蛋白表达明显上调;细胞体外侵袭能力亦显著下降。结论 S100A4能通过调控MMP-2和E-cadherin的表达影响胃癌细胞MGC-803的侵袭能力。
Objective To explore the effects of si RNA-mediated S100A4 silencing on MMP-2 and E-cadherin expression and invasive ability of human gastric cancer cell lines MGC-803.Methods The S100A4-si RNA was transfected into human gastric cancer MGC-803 cells.The expressions of S100 A4 m RNA and protein were detected by Real-time RT-PCR and Western blot to select S100 A4 silencing and negative control cells.The levels of MMP-2 and Ecadherin proteins were determined by Western blot.The changes of invasive abilities were detected by Transwell chamber model.Results The human gastric cancer MGC-803 cells of S100 A4 silencing were successfully established.Compared with the control group,the levels of S100 A4 gene and protein were significantly decreased,the expression levels of MMP-2 protein decreased and the level of E-cadherin increased significantly,the invasive ability was reduced obviously In MGC-803 cells of S100 A silencing.Conclusion S100 A4 could regulate the expression of MMP-2 and E-cadherin to affect the invasive abilities of human gastric cancer MGC-803 cells.
作者
赵滢
高立红
王楠
鲍晨辉
王文学
张天彪
ZHAO Ying;GAO Li-hong;WANG Nan;BAO Chen-hui;WANG Wen-xue;ZHANG Tian-biao(Department of Gastrointestinal Nutrition Surgery,Shengjing Hospital of China Medical University,Liaoning Shenyang 110004;State key Laboratory of Robotics,Shenyang Institute of Automation,Chinese Academy of Sciences;Department of Biochemistry and Molecular Biology,College of Basic Medical,China Medical University,Liaoning Shenyang 110122,China)
出处
《解剖科学进展》
2018年第4期364-367,共4页
Progress of Anatomical Sciences
基金
国家自然科学青年基金(61703435)
辽宁省博士科研启动基金(201601122)
机器人学国家重点实验室开放基金(2013225021)
