【作者】 檀碧波； 李勇； 赵群； 范立侨； 刘羽； 李兆星； 安昭杰；

【Author】 TAN Bi-bo;LI Yong;ZHAO Qun;FAN Li-qiao;LIU Yu;LI Zhao-xing;AN Zhao-jie;Department of General Surgery,the Fourth Affiliated Hospital,Hebei Medical University;

【机构】 河北医科大学第四医院外三科；

【摘要】 目的应用RNA干扰技术抑制胃癌细胞Vav3表达,检测转染前后胃癌细胞株体外化疗药敏性及凋亡相关蛋白的影响,对相关机制进行探讨。方法 QRTPCR及蛋白印迹技术检测Vav3在胃癌、癌旁组织、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;检测转染前后凋亡抑制蛋白家族xIAP、Survivin、Livin及Caspase-3、Caspase-8的表达。结果①Vav3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(均P<0.01);②Vav3-siRNA转染SGC7901后Vav3表达明显受到抑制(P<0.01);③Vav3-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(均P<0.05);④转染后SGC7901中xIAP、Survivin的表达均较转染前降低(均P<0.05),Livin表达在转染前后无明显变化(P>0.05);Caspase-3、Caspase-8表达及活性升高(均P<0.05)。结论 Vav3可通过调控胃癌细胞的凋亡抑制途径而参与胃癌多药耐药,抑制Vav3表达可逆转胃癌耐药细胞的多药耐药性,该作用与Vav3能调节部分凋亡抑制蛋白家族成员的表达有关。更多还原

【Abstract】 Objective In present study,Vav3 gene was inhibited with RNA interference technology,then chemosensitivity in vitro of gastric cancer cells,exprssion of inhibitor of apoptosis proteins(IAPs) were tested,and mechanism was explored.Methods The expressions of Vav3 in gastric cancer tissus,tumor-adjacent tissues,human gastric cancer cell line SGC7901,gastric epithelial cell line GES-1 were tested by Western blot.Then Vav3-siRNA was synthesized and tansfected into SGC7901.Inhibition rates to tumor cells by chemotherapeutic agents(5-FU,L-OHP) were measured with MTT assay in vitro.The expressions of xIAP,Survivin,Livin and Caspase-3,Caspase-8 were also determined.Results Overexpressions of Vav3 were detected in gastric cancer tissues and gastric cell line(both P<0.01).Expression of Vav3 was effectively inhibited by Vav3-siRNA(P<0.01).Inhibition rates to tumor cells by 5-FU,L-OHP increased after Vav3-siRNA tansfected into gastric cancer cells after 48 h(both P<0.05).The expressions of xIAP,Survivin were lower in cells after Vav3-siRNA tansfected(both P<0.05),and no obviously change was found in Livin(P>0.05);expression and activity of Caspase-3,Caspase-8 increased after Vav3-siRNA tansfected into SGC7901(all P<0.05).Conclusions Vav3 was invovled in multidrug resistance(MDR)of gastric cancer by regulating apoptotic pathways,and inhibiting Vav3 can reverse MDR of gastric cancer cells by regulating some IAPs.更多还原