MAPK/ERK通路及泛素连接酶c-Cbl调控埃克替尼抑制肺癌eCC827细胞增殖和诱导细胞凋亡的研究

摘要

Objective:To investigate the role of c - Cbl involved in inactivation of MAPK/ ERK Pathway induced by Icotinib. Methods:The effect of Icotinib on the Proliferation of eCC827 was detected by MTT. Cell aPoPtosis was determined by flow cytometry with Annexin V - PI staining. Protein exPression levels were detected by Western blot. All exPerimental data was analyzed with SPSS 18. 0 software. Results:The IC50 value of eCC827 incuabted with Ico-tinib for 24h was 155. 6 μmol/ L;Icotinib induced the aPoPtosis of eCC827 in a dose - dePendent manner. Western blot results showed that Icotinib decreased the exPression level of P - EGFR,P - ERK and c - Cbl. Conclusion:Ico-tinib significantly inhibits eCC827 Proliferation by inducing aPoPtosis. The reason is that c - Cbl leads to inactivation of MAPK/ ERK Pathway as showed by downregualtion of P - EGFR and P - ERK.%目的：研究 MAPK/ ERK 通路及泛素连接酶 c - Cbl 调控埃克替尼抑制 eCC827细胞增殖和诱导细胞凋亡的作用机制。方法：采用 MTT 法检测 eCC827对埃克替尼的敏感性，Annexin V - PI 流式细胞术检测细胞凋亡，Western blot 检测相关蛋白表达，SPSS 18.0进行统计学分析。结果：埃克替尼处理 eCC827细胞24h 的IC50为155.6μmol/ L。埃克替尼诱导 eCC827细胞早期凋亡并具有剂量依赖性。Western blot 结果显示埃克替尼可诱导 P - EGFR 和 P - ERK 下调并引起 c - Cbl 下调。结论：埃克替尼能够明显抑制 eCC827细胞增殖，诱导细胞早期凋亡。其机制可能与 c - Cbl 参与的 P - EGFR 和 P - ERK 蛋白表达水平下调，从而抑制细胞增殖通路活化有关。