2017 年第 2 期 第 15 卷

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

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关键词：克唑替尼AKT信号通路EML4-ALK阳性肺癌凋亡迁移

• 摘要：
• 【摘要】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试验检测20、40、80、160、320、640nmol/L的克唑替尼作用于细胞48h后对细胞增殖的影响，计算IC50；300nmol/L克唑替尼处理细胞24、48、72h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 克唑替尼能显著抑制人肺癌细胞株H2228增殖，具有浓度依赖性（P<0.01），根据IC50值选择300nmol/L克唑替尼为研究对象；克唑替尼能显著促进细胞凋亡，抑制细胞迁移，具有时间依赖性（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，且具有时间依赖性（P<0.01）。AKT蛋白表达在各个时间差异不显著（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。
• Objective To investigate the role and mechanism of AKT signaling pathway in the apoptosis and migration of EML4-ALK positive lung cancer cells.Methods Human lung cancer cell line H2228 were cultured, after 20,40, 80, 160, 320, 640nmol/L of crizotinib treated cells for 48h, cell proliferation was detected by CCK8 test, and calculation IC50; 300nmol/L crizotinib treated cells 24, 48, 72h, cell apoptosis was detected by flow cytometry. Transwell was used to examine cell migration; the expression of Bcl-xL, Bcl-2, Bim, AKT, p-AKT protein was detected by Western blot. Results Crizotinib can significantly inhibit the proliferation of human lung cancer cell line H2228 in a concentration dependent manner (P<0.01), according to the IC50 value 300nmol/L crizotinib was selected as the research object; crizotinib can significantly promote cell apoptosis, inhibit cell migration with time dependent (P<0.01); crizotinib can down Bcl-xL, Bcl-2, and p-AKT protein expression, up Bim protein expression with time dependent (P<0.01). AKT protein expression was not significant at all time (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.