维拉帕米通过调节Bcl-2表达对胃癌细胞株耐阿霉素的影响  

Bcl-2 Targeted by Verapamil in the Prevention of Drug Resistance to Adriamycin in Gastric Cancer Cells

作　　者：滕小路 刘淼 张腾跃[2] 黄金[2] 王成[2] 杨国红 范平生[2] TENG Xiao-lu;LIU Miao;ZHANG Teng-yue(Xinxiang Medical University,Xinxiang 453000,China;Anhui Provincial Cancer Hospital, Hefei 230001, China)

机构地区：[1]新乡医学院,河南新乡453000 [2]安徽省肿瘤医院,安徽合肥230001

出　　处：《肿瘤学杂志》2018年第4期328-335,共8页Journal of Chinese Oncology

基　　金：国家自然科学基金(81350005);安徽省科技厅2012年度重点科研计划项目(12070403058);安徽省自然科学基金(1408085MH211);安徽省“十二五”临床重点专科建设项目;安徽省自然科学基金(1708085MH228)

摘　　要：[目的]探讨Bcl-2在维拉帕米逆转胃癌阿霉素化疗耐药中的作用及初步机制研究。[方法 ]MTT法检测维拉帕米逆转胃癌细胞系（SGC-7901、BGC-823、AGS、HGC-27）对3种化疗药物（氟尿嘧啶、阿霉素、顺铂）的耐药能力;流式细胞仪Annexin V-FITC/PI法检测维拉帕米对阿霉素作用下的胃癌细胞系（SGC-7901、HGC-27）凋亡水平的影响。Western blotting检测Bcl-2蛋白表达水平。[结果 ]SGC-7901细胞系在单药阿霉素组和阿霉素联合维拉帕米组中的半数抑制浓度（IC50）分别为1.51±0.12μg/ml、0.22±0.01μg/ml,两组差异有统计学意义（P〈0.05）;HGC-27细胞中分别为1.24±0.19μg/ml、1.06±0.08μg/ml,两组差异无统计学意义（P〉0.05）。凋亡实验表明在SGC-7901细胞系中,单药阿霉素组中的细胞凋亡率为36.76%,阿霉素联合维拉帕米组为61.10%,两组比较差异有统计学意义（P〈0.05）。HGC-27细胞系中,阿霉素组细胞凋亡率19.79%,阿霉素联合维拉帕米组为21.92%,两组比较差异无统计学意义（P〉0.05）。SGC-7901细胞中维拉帕米促进ADM组胃癌细胞凋亡指数明显高于HGC-27（P〈0.05）。q RT-PCR实验显示Bcl-2可能介导维拉帕米促进胃癌细胞凋亡;Western blotting结果显示在SGC-7901细胞中,阿霉素联合维拉帕米组Bcl-2的表达明显低于单药阿霉素组,与对照组比较差异有统计学意义（P〈0.05）;而HGC-27中2组间差异无统计学意义。[结论]维拉帕米通过下调Bcl-2表达增强维拉帕米逆转胃癌细胞化疗耐受的能力,并促进胃癌细胞凋亡。[Objective]To investigate the chemotherapy resistance reversal in gastric cancer cells with verapamil,screen the target gene Bcl-2 associated with the above reversal process and study its mechanism. [Methods] The ability of verapamil in reversing the chemotherapy resistance of fluorouracil,doxorubicin and cisplatin in gastric cancer cell lines SGC-7901,BGC-823,AGS and HGC-27 was detected with the CCK-8 method. The effect of verapamil on the apoptosis of doxorubicin-based gastric cancer cell lines（SGC-7901,HGC-27） was also tested with the Annexin VFITC/PI method. The protein expression levels of Bcl-2 were detected with Western blotting.[Results] The IC50 values of SGC-7901 cell lines in ADM（adriamycin） group and ADM-VRP（adriamycin-verapamil） combination group were 1.51±0.12μg/ml and 0.22±0.01μg/ml,respectively,there was an obvious difference between the two groups（P〈0.05）. The IC50 values of HGC-27 cells were 1.24±0.19μg/ml and 1.06±0.08μg/ml,there was no obvious difference between the two groups（P〈0.05）. The apoptosis experiment showed that apoptosis rates of the SGC-7901 cell line in ADM group and ADM-VRP combination group were 36.76% and 61.10%,respectively. The data were significantly different from that of the blank group（P〈0.05）. Apoptosis rates of the HGC-27 cell line in ADM group and ADM-VRP combination group were 19.79% and 21.92%,respectively. The apoptosis index of the SGC-7901 cells promoted by verapamil in ADM group was significantly higher than that of HGC-27 cells,and there was a significant difference between them（P〈0.05）. A q RT-PCR experiment results showed that Bcl-2 might mediate verapamil in promoting the apoptosis of gastric cancer cells. Western blotting results showed that the Bcl-2 expression in SGC-7901 cells treated with the combination of adriamycin and verapamil was significantly lower than that treated with adriamycin alone（P〈0.05）. The data had significant differences with the control group while the differences among the four HGC-27

关 键 词：胃肿瘤 维拉帕米 多柔比星 耐药性 BCL-2 

分 类 号：R735.2[医药卫生—肿瘤]