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目的探讨阿法替尼对体外人卵巢癌A2780及A2780T细胞增殖的影响及其可能的作用机制。方法不同浓度的阿法替尼作用于体外培养的人卵巢癌A2780及A2780T细胞48 h,采用四甲基偶氮唑蓝比色法(MTT)检测阿法替尼对卵巢癌A2780及A2780T细胞增殖的影响;采用Bliss法计算半数抑制浓度(IC50);采用Western Blot法检测阿法替尼对卵巢癌A2780T细胞中EGFR、p-EGFR、HER-2、p-HER-2、AKT及p-AKT等相关蛋白表达的影响。结果阿法替尼可浓度依赖性的抑制人卵巢癌A2780及A2780T细胞的体外增殖,IC50分别为3.28μmol·L-1及6.81μmol·L-1;阿法替尼可抑制卵巢癌A2780T细胞中表皮生长因子受体(EGFR)、人表皮生长因子受体2(HER-2)及蛋白激酶B(AKT)的磷酸化,而对EGFR、HER-2及AKT的总蛋白表达水平没有影响。结论阿法替尼可通过抑制EGFR通路的激活,下调EGFR下游的PI3K/AKT通路,进而抑制人卵巢癌细胞的体外增殖,是一种有开发前景的卵巢癌化疗药物。
Objective To investigate the effects of afatinib on the proliferation of human ovarian cancer A2780 and A2780T cells in vitro and its possible mechanism. Methods Afatinib was administered to human ovarian cancer A2780 and A2780T cells in vitro for 48 h. The proliferation of ovarian cancer A2780 and A2780 T cells was detected by MTT assay. The half-inhibitory concentration (IC50) was calculated by Bliss method. The expression of EGFR, p-EGFR, HER-2, p-HER-2, AKT and p-AKT was detected by Western Blot in the ovarian cancer A2780T cells And other related protein expression. Results Afatinib could inhibit the proliferation of A2780 and A2780T cells in vitro in a concentration-dependent manner with IC50 of 3.28μmol·L-1 and 6.81μmol·L-1, respectively. Afatinib could inhibit the proliferation of A2780T cells Epidermal growth factor receptor (EGFR), human epidermal growth factor receptor 2 (HER-2) and protein kinase B (AKT) phosphorylation, but had no effect on the total protein expression level of EGFR, HER-2 and AKT. Conclusion Afatinib can inhibit the activation of EGFR pathway and down-regulate the PI3K / AKT pathway downstream of EGFR, thereby inhibiting the proliferation of human ovarian cancer cells in vitro. It is a promising chemotherapeutic agent for ovarian cancer.