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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 investigate the role of Vav3 in multidrug resistance (MDR) in gastric cancer and its possible mechanism. Methods The expression of Vav3 in gastric cancer tissues, adjacent non-cancerous tissues and human gastric cancer cell lines SGC7901 and gastric epithelial cells GES-1 was detected by quantitative reverse transcriptase-polymerase chain reaction (QRT-PCR) and Western blotting. The siRNA against Vav3 was synthesized, And then transfected into SGC7901 cells. The inhibitory rates of 5-fluorouracil (5-FU) and oxaliplatin (L-OHP) on the gastric cancer cells before and after transfection were detected by MTT assay. Apoptosis was inhibited by fluorescence quantitative RT-PCR and Western blotting Protein family members xIAP, Survivin, Livin expression, and detection Caspase-3, Caspase-8 expression and activity. Results The expression of Vav3 in gastric cancer tissues and cell lines was higher than that in paracancerous tissues and gastric epithelial cell lines (P <0.05). The expression of Vav3 in Vav3-siRNA transfected SGC7901 cells was significantly inhibited (P <0.01). Vav3 siRNA transfected SGC7901 After 48h, the inhibitory effect of 5-FU and L-OHP on tumor cells were significantly increased (P <0.05). The expression of xIAP and Survivin in transfected SGC7901 cells was lower than that before transfection (P <0.05) There was no obvious change before and after dyeing. The expression and activity of Caspase-3, Caspase-8 after transfection increased (P <0.05). Conclusion Vav3 may be involved in the multidrug resistance of gastric cancer by regulating the apoptosis inhibitory pathway of gastric cancer cells. Inhibiting Vav3 expression may reverse the chemoresistance of gastric cancer cells.