阿法替尼增强耐药性卵巢癌细胞对多柔比星化疗敏感性的机制研究

Mechanism of afatinib enhancing the chemosensitivity of drug-resistant ovarian cancer cells to adriamycin

目的探讨阿法替尼增强耐药性卵巢癌细胞对多柔比星化疗敏感性的机制。方法选取人卵巢癌耐药细胞株A2780T,采用噻唑蓝(MTT)法检测不同浓度(0.63、1.25、2.50、5.00μmol/L)阿法替尼对A2780T细胞增殖的影响。分别将多柔比星、多柔比星+0.63μmol/L阿法替尼、多柔比星+1.25μmol/L阿法替尼、多柔比星+2.50μmol/L阿法替尼处理的A2780T细胞作为多柔比星组、A组、B组、C组,比较4组细胞的半数抑制浓度。采用蛋白质印迹法(Western blot)检测4组细胞中表皮生长因子受体(EGFR)、磷酸化EGFR(p-EGFR)、人表皮生长因子受体2(HER2)、磷酸化HER2(p-HER2)及三磷酸腺苷结合盒转运子B亚家族成员1(ABCB1)的相对表达量。结果阿法替尼抑制A2780T细胞增殖呈浓度依赖性,0.63、1.25、2.50、5.00μmol/L阿法替尼处理的A2780T细胞的增殖抑制率分别为(2.30±0.82)%、(4.50±1.12)%、(8.10±1.43)%和(24.49±3.22)%。A组、B组、C组A2780T细胞的半数抑制浓度均低于多柔比星组,B组、C组A2780T细胞的半数抑制浓度均低于A组,C组A2780T细胞的半数抑制浓度低于B组,差异均有统计学意义(P﹤0.05)。A组、B组、C组A2780T细胞中p-EGFR、p-HER2、ABCB1蛋白的相对表达量均低于多柔比星组,B组、C组A2780T细胞中p-EGFR、p-HER2、ABCB1蛋白的相对表达量均低于A组,C组A2780T细胞中p-EGFR、p-HER2、ABCB1蛋白的相对表达量均低于B组,差异均有统计学意义(P﹤0.05)。结论阿法替尼可增强耐药性卵巢癌细胞A2780T对多柔比星的化疗敏感性,可能与阿法替尼下调p-EGFR、p-HER2和ABCB1蛋白表达有关。

Objective To investigate the mechanism of afatinib enhancing the chemosensitivity of drug-resistant ovarian cancer cells to adriamycin.Method The human ovarian cancer drug-resistant cell line A2780T was selected,and the methyl thiazolyl tetrazolium(MTT)method was used to detect the effect of different concentrations(0.63,1.25,2.50,5.00μmol/L)of afatinib on the proliferation of A2780T cells.Treatment regimens including doxorubicin,doxorubicin+0.63μmol/L afatinib,doxorubicin+1.25μmol/L afatinib,and doxorubicin+2.50μmol/L afatinib were used on these cells and divided into doxorubicin group,group A,group B,and group C.The half maximal inhibitory concentration(IC50)of the four groups of cells was compared.The expression of epidermal growth factor receptor(EGFR),phosphorylated-EGFR(p-EGFR),human epidermal growth factor receptor 2(HER2),phosphorylated-HER2(p-HER2),and ATP binding cassette subfamily B member 1(ABCB1)were measured by Western blot.Result Afatinib inhibited A2780T cell proliferation in a concentration-dependent manner,the inhibition rates of A2780T cells cultured in 0.63,1.25,2.50 and 5.00μmol/L afatinib were(2.30±0.82)%,(4.50±1.12)%,(8.10±1.43)%and(24.49±3.22)%,respectively.The IC50 of A2780T cells in group A,B and C were lower than that of doxorubicin group,the IC50 of A2780T cells in group B and C were lower than that of group A,and the IC50 of A2780T cells in group C was lower than that of group B,the differences were statistically significant(P<0.05).The relative expression levels of p-EGFR,p-HER2 and ABCB1 protein in A2780T cells of group A,B and C were lower than those of the doxorubicin group,the relative expression levels of p-EGFR,p-HER2 and ABCB1 protein in A2780T cells of group B and C were lower than those of group A,and the relative expression levels of p-EGFR,p-HER2 and ABCB1 protein in A2780T cells of group C were lower than those of group B,and the differences were statistically significant(P<0.05).Conclusion Afatinib can enhance the chemosensitivity of drug-resistant ovarian cancer cell A2780T to adriamycin,which may be related to the down-regulatory effect of afatinib on p-EGFR,p-HER2,and ABCB1.