AKT信号通路在克唑替尼诱导的EML4-ALK阳性肺癌细胞凋亡迁移中的作用及机制研究

目的 探讨AKT信号通路在克唑替尼诱导的EML4-ALK阳性肺癌细胞凋亡迁移中的作用及机制研究.方法 培养人肺癌细胞株H2228,CCK8实验检测20、40、80、160、320、640 nmol/L的克唑替尼作用于细胞48 h后对细胞增殖的影响,计算半抑制浓度(IC50);300 nmol/L克唑替尼处理细胞24、48、72 h后,流式细胞仪检测细胞凋亡率,Transwell小室检测细胞迁移数;Western Blot检测Bcl-xL、Bcl-2、Bim、AKT、p-AKT蛋白表达.结果 克唑替尼能明显抑制人肺癌细胞株H2228增殖(P﹤0.01),根据IC50值选择300 nmol/L克唑替尼为研究对象;克唑替尼能明显促进细胞凋亡,抑制细胞迁移(P﹤0.01);克唑替尼能下调Bcl-xL、Bcl-2、p-AKT蛋白表达,上调Bim蛋白表达(P﹤0.01).AKT蛋白表达水平在克唑替尼作用的各个时间点间比较,差异无统计学意义(P﹥0.05).结论 克唑替尼通过AKT信号通路抑制人肺癌细胞株H2228迁移,并通过调节Bcl-xL、Bcl-2、Bim蛋白表达促进细胞凋亡.

The role and mechanism of AKT signaling pathway in the apoptosis and migration of EML4-ALK positive lung cancer cells

Objective To investigate the role and mechanism of AKT signaling pathway in the apoptosis and migra-tion of EML4-ALK positive lung cancer cells. Method Human lung cancer cell line H2228 were cultured, and after treatment of 20, 40, 80, 160, 320, 640 nmol/L of crizotinib for 48 h, the cell proliferation was detected by CCK8 assay, and IC50 was calculated;besides, 300 nmol/L crizotinib was used to treat cells for 24, 48, 72 h, after which the cell apopto-sis was detected by flow cytometry. Transwell chamber was used to examine cell migration; the expression of Bcl-xL, Bcl-2, Bim, AKT, p-AKT protein was detected by Western Blot. Result Crizotinib significantly inhibited the prolifera-tion of human lung cancer cell line H2228 in a concentration dependent manner (P<0.01), according to the IC50 value, 300 nmol/L crizotinib was selected as the target concentration;crizotinib significantly promoted cell apoptosis, and inhib-ited cell migration (P<0.01);meanwhile, it can reduce the level of Bcl-xL, Bcl-2, and p-AKT protein expression, while the Bim protein expression was up-regulated (P<0.01). AKT protein expression in all time points of the treatment with crizotinib were of no significant difference (P>0.05). Conclusion Crizotinib inhibits the migration of human lung cancer cell line H2228, and promotes cell apoptosis through regulating the expression of Bcl-xL, Bcl-2 and Bim protein by the AKT signaling pathway.