Evi1基因对卵巢癌化疗敏感性的影响机制分析

Effect mechanism of Evi1 gene on chemotherapy sensitivity of ovarian cancer

目的分析异位病毒整合位点1(Evi1)基因影响卵巢癌化疗敏感性的机制。方法选取正常培养卵巢癌细胞系A2780和SKOV3细胞,对照组不进行siRNA技术干扰,干扰组采用siRNA技术干扰Evi1基因的表达。采用四甲基偶氮唑盐比色法(MTT法)检测2组细胞的半数抑制浓度(IC50);原位末端转移酶标记法(TUNEL法)检测2组卵巢癌细胞的凋亡情况;蛋白免疫印迹法(Western Blotting)检测磷脂酰肌醇-3-激酶(PI3K)/丝/苏氨酸蛋白激酶(AKT)/m TOR信号通路、多药耐药相关蛋白1(MRP1)、P糖蛋白(P-gp)蛋白的表达。结果顺铂对A2780和SKOV3细胞增殖的抑制作用呈浓度依赖性;与对照组相比,干扰组A2780细胞和SKOV3细胞Evi1蛋白和mRNA表达量显著下降(P<0.05);干扰组顺铂对A2780细胞和SKOV3细胞的IC50显著下降(P<0.05),细胞凋亡率显著升高(P<0.05);pPI3K、p-AKT、p-m TOR、MRP1和P-gp蛋白的表达量显著下调,差异有统计学意义(P<0.05)。结论Evi1基因可能通过抑制PI3K/AKT/m TOR信号通路,下调耐药相关蛋白的表达,细胞凋亡率升高,降低细胞耐药性的产生,提高化疗敏感性。

Objective To analyze the mechanism of ectopic viral integration site-1(Evi1)gene affecting chemotherapy sensitivity of ovarian cancer.Methods The normal cultured ovarian cancer cell lines A2780 and SKOV3 cells were selected.The control group was not given interfere with siRNA technology.The siRNA technology was performed to interfere Evi1 gene expression in interference group.The half maximal inhibitory concentration(IC50)of both groups was detected by tetramethylazolium salt colorimetric method(MTT method).The apoptosis of ovarian cancer cells of both groups was detected by TUNEL.Phosphatei-dylinositol 3 kinase(PI3 K)/serine protein kinase(AKT)/AKT/m TOR signaling pathway,expression of multidrug resistance associated proteins 1(MRP1)and P-glycolprotein(P-gp)were detected by Western Blotting.Results The inhibitory effect of cisplatin on proliferation of A2780 and SKOV3 cells showed concentration-dependence.Compared with control group,the expression quantities of Evi1 protein and mRNA in A2780 cells and SKOV3 cells were significantly decreased in interference group(P<0.05),IC50 of A2780 cells and SKOV3 cells was significantly decreased(P<0.05),while apoptosis rate was significantly increased(P<0.05).The expression quantities of p-PI3 K,p-AKT,p-m TOR,MRP1 and P-gp were significantly down-regulated(P<0.05).Conclusion The Evi1 gene may inhibit the PI3 K/AKT/m TOR signaling pathway,down-regulate the expression of resistance-related proteins,increase the rate of cell apoptosis,reduce the generation of cell drug resistance,and improve chemotherapy sensitivity.