抑制Vav3基因表达对胃癌细胞凋亡抑制蛋白及肿瘤多药耐药性的影响  

Effect of Vav3 gene inhibition on inhibitor of apoptosis proteins and multidrug resistance in gastric cancer cells

作　　者：檀碧波[1] 李勇[1] 范立侨[1] 赵群[1] 王冬[1] 刘羽[1] 

机构地区：[1]河北医科大学第四医院外三科,石家庄050011

出　　处：《第二军医大学学报》2015年第2期136-141,共6页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(81072033;81372580);河北省自然科学基金(C2010000619);河北省普通高校强势特色学科资助项目(冀教高[2005]52);河北省科技支撑项目(14277779D);河北省卫生厅重大医学科研课题(zd2013040)~~

摘　　要：目的探讨Vav3基因在胃癌多药耐药性(MDR)中的作用及可能机制。方法采用荧光定量反转录聚合酶链(QRT-PCR)及蛋白印迹技术检测Vav3在胃癌、癌旁组织及人胃癌细胞株SGC7901、胃上皮细胞GES-1中的表达;合成针对Vav3的siRNA,并转染SGC7901;MTT法检测氟尿嘧啶(5-FU)、奥沙利铂(L-OHP)对转染前后胃癌细胞的抑制率;荧光定量RT-PCR和蛋白质印迹法检测转染前后凋亡抑制蛋白家族成员xIAP、Survivin、Livin表达,并检测Caspase-3、Caspase-8表达及活性。结果 Vav3在胃癌组织及细胞株的表达高于癌旁组织及胃上皮细胞株(P<0.05);Vav3-siRNA转染SGC7901后Vav3表达明显受到抑制(P<0.01);Vav3-siRNA转染SGC7901 48h后5-FU、L-OHP对肿瘤细胞的抑制作用均明显增强(P<0.05);转染后SGC7901中xIAP、Survivin的表达均较转染前降低(P<0.05),Livin表达在转染前后无明显变化;转染后Caspase-3、Caspase-8表达及活性升高(P<0.05)。结论 Vav3可通过调控胃癌细胞凋亡抑制途径参与胃癌多药耐药,抑制Vav3表达可能有助于逆转胃癌细胞的化疗耐药。Objective To explore the role of Vav3 gene in multidrug resistance（MDR）of gastric cancer and the related mechanism.Methods QRT-PCR was used to examine the expressions of Vav3 gene in gastric cancer tissues,tumor-adjacent tissues,human gastric cancer cell line SGC7901,and gastric epithelial cell line GES-1.Then Vav3-siRNA was synthesized and tansfected into SGC7901 cells. MTT assay was then used to determine the inhibition rates of tumor cells exposed to chemotherapeutic agents（5-FU,L-OHP）before and after Vav3-siRNA transfection.Real-time RT-PCR and Western blotting analysis were used to observe the expressions of inhibitor of apoptosis proteins（IAPs）：xIAP,Survivin,and Livin;meanwhile,the expression and activity of Caspase-3and Caspase-8 were also determined.Results Vav3 was over-expressed in gastric cancer tissues and gastric cell line compared with those in tumor-adjacent tissues and gastric epithelial cell line GES-1（P〈0.05）.Expression of Vav3 was significantly inhibited by Vav3-siRNA（P〈0.01）.Inhibition rates of tumor cells exposed to 5-FU and L-OHP were significantly increased 48 hafter Vav3-siRNA tansfection（P〈0.05）.The expressions of xIAP and Survivin were significantly decreased in cancer cells after Vav3-siRNA tansfection（bothP〈0.05）,and no notable change was found for Livin expression;also the expression and activity of Caspase-3and Caspase-8protein were significantly increased after Vav3-siRNA tansfection in SGC7901cells（allP〈0.05）.Conclusion Vav3 can participate in MDR of gastric cancer by regulating apoptotic pathways,and inhibition of Vav3 can help reverse MDR of gastric cancer cells by regulating some IAPs.

关 键 词：胃肿瘤 Vav3 多药抗药性 化疗敏感性 凋亡抑制蛋白质类 

分 类 号：R735.2[医药卫生—肿瘤]