ABCG2基因在非小细胞肺癌细胞耐药中的作用和机制探究

Expression of ABCG2 gene in multidrug-resistant non-small cell lung cancer cells and its possible mechanism

目的:分析ABCG2基因在多耐药非小细胞肺癌(non small cell lung cancer,NSCLC)细胞的表达变化及其可能参与的机制。方法:选择肺癌细胞A549及耐药A549进行细胞培养,分别给予顺铂及依托泊苷化疗药物治疗;取耐药A549细胞分为空白组(不处理)、对照组(给予siRNA对照)及siRNA-ABCG2组(给予siRNA-ABCG2)。采用CCK8法检测各组细胞的药物抑制率,采用Western blot法检测3组细胞中ABCG2蛋白表达情况。结果:2组细胞药物抑制率变化呈剂量依赖性,同剂量中耐药A549组药物抑制率明显低于A549组。耐药A549组细胞ABCG2蛋白表达水平明显高于A549组。3组细胞药物抑制率呈剂量依赖性,其中同剂量的siRNA-ABCG2组药物抑制率明显高于空白组及对照组。siRNA-ABCG2组细胞ABCG2、p-AKT、p-mTOR蛋白表达水平明显低于空白组及对照组,分别使用p-AKT、p-mTOR的激活剂后,可显著降低干扰ABCG2基因后导致的顺铂和依托泊苷抑制率的增加。结论:ABCG2通过调控PI3K/AKT信号通路在耐药A549的耐药性中发挥重要作用,沉默ABCG2基因可改善耐药A549细胞的耐药性,提示ABCG2可作为改善NSCLC细胞对顺铂及依托泊苷化疗药物的耐药性的潜在靶点。

OBJECTIVE To analyze the expression of ABCG2 gene in non small cell lung cancer(NSCLC) cells and its possible mechanism.METHODS Lung cancer cells A549 and drug-resistant A549 were selected for cell culture,and cisplatin and etoposide chemotherapy drugs were treated respectively;drug-resistant A549 cells were divided into blank group,control group and siRNA-ABCG2 group.The blank group was not treated,the control group was given siRNA control,and siRNA-ABCG2 group was given siRNA-ABCG2.CCK8 method was used the drug inhibition rate of cells in each group was detected,and the expression of ABCG2 protein in the three groups was detected by Western blot.RESULTS The drug inhibition rates of the two groups were dose-dependent on cisplatin and etoposide.The drug inhibition rate of drug-resistant A549 group was significantly lower than that of A549 group at the same dose.The expression level of ABCG2 protein in drug-resistant A549 group was significantly higher than that in A549 group.The drug inhibition rates of the three groups were dose-dependent on cisplatin and etoposide.The drug inhibition rates of siRNA-ABCG2 group at the same dose were significantly higher than those of blank group and control group.The expression levels of ABCG2 and p-AKT protein in siRNA-ABCG2 group were significantly lower than those in blank group and control group.CONCLUSION ABCG2 plays an important role in drug resistance of drug-resistant A549 by regulating PI3 K/AKT signaling pathway.Silencing ABCG2 gene can improve the drug resistance of drug-resistant A549 cells,suggesting that ABCG2 can be used as a potential target for improving the drug resistance of NSCLC cells to cisplatin andetoposide chemotherapy drugs.