自噬调节在结肠癌细胞化疗耐药性的作用机制研究

目的:探究自噬调节在结肠癌细胞化疗过程中耐药性的作用机制.方法:以顺铂诱导人结肠癌细胞系EC9706细胞自噬,观察自噬抑制剂3-MA对EC9706细胞的影响.CCK8法测定细胞生长情况.流式细胞术检测细胞凋亡和细胞周期.MDC检测自噬.Western blot法检测Beclin-1、PI3K Ⅲ、LC3-Ⅰ、LC3-Ⅱ...

Mechanism of autophagy regulating chemoresistance in colon cancer cells

Objective:To explore the mechanism of autophagy regulation in drug resistance of colon cancer cells during chemotherapy.Methods:Autophagy of human colon cancer cell line EC9706 was induced by cisplatin,and the effect of autophagy inhibitor 3-MA on EC9706 cells was observed.The cell growth was determined by CCK8 method.Cell apoptosis and cell cycle were detected by flow cytometry.MDC detects autophagy.Western blot method to detect Beclin 1,PI3KⅢ,LC3-Ⅰ,LC3-Ⅱ protein expression.Results:The inhibition rate of EC9706 cells in combination group was(57.34±0.55)%,which was significantly higher than that in cisplatin group(32.00±0.32)%.The inhibitory rate of EC9706 cells in combination group and cisplatin group was significantly higher than that in control group(5.22±0.06)%.The number of autophagy vacuoles in MDC staining of cisplatin group was(1.72±0.12),which was significantly higher than that in control group(1.00±0.06)and combined group(0.99±0.35).The apoptosis rate of EC9706 cells in the combined group was(29.12±0.34)%,significantly higher than that in the cisplatin group(19.76±0.15)%,and the apoptosis rate of cells in the G0/G1 and G2/M stages were(54.68±0.35)% and(2.33±0.05)%,respectively,which were significantly lower than that in the cisplatin group(58.45±0.12)% and(7.56±0.16)%,respectively.The apoptosis rate of cells in the S stage was(44.78±0.45)%,significantly higher than that in the cisplatin group(38.25±0.65)%(P<0.05).Joint and cisplatin group Beclin 1,PI3KⅢ,LC3-Ⅰ,multiples LC3-Ⅱ protein expression were significantly higher than that of control group,and the joint group of Beclin 1,PI3KⅢ,LC3-Ⅰ,multiples LC3-Ⅱ protein expression were significantly lower than that of cisplatin group(P<0.05).Conclusion:Autophagy may be a self-protection mechanism of cisplatin induced colon cancer cells,and inhibition of autophagy may be a new strategy of adjuvant chemotherapy for colon cancer cells.