| 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 48 h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。 |
| 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 |
| DOI：10.3724/SP.J.1008.2015.00136 |
| 投稿时间：2014-06-24修订日期：2014-08-15 |
| 基金项目: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). |
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| Effect of Vav3 gene inhibition on inhibitor of apoptosis proteins and multidrug resistance in gastric cancer cells |
| TAN Bi-bo,LI Yong*,FAN Li-qiao,ZHAO Qun,WANG Dong,LIU Yu |
(The Third Department of Surgery, the Fourth Affiliated Hospital of Hebei Medical University, Shijiazhuang 050011, Hebei, China
*Corresponding author) |
| Abstract: |
| 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-3 and 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 h after Vav3-siRNA tansfection (P<0.05). The expressions of xIAP and Survivin were significantly decreased in cancer cells after Vav3-siRNA tansfection (both P<0.05), and no notable change was found for Livin expression; also the expression and activity of Caspase-3 and Caspase-8 protein were significantly increased after Vav3-siRNA tansfection in SGC7901 cells (all P<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. |
| Key words: stomach neoplasms Vav3 multiple drug resistance chemosensitivity inhibitor of apoptosis proteins |
