自噬在顺铂诱导的食管癌细胞死亡中的作用

目的 观察自噬在食管癌EC9706细胞顺铂处理过程中的表达,探讨其表达与细胞死亡的关系.方法 对数生长期EC9706细胞与顺铂共同作用0、3、6、12、24 h后,Western blot法检测微管相关蛋白轻链3(LC3)的表达;自噬特异性阻断剂3-甲基腺嘌呤(3-MA)分别在顺铂前1 h、后1 h加入共同作用24 h,用CCK-8法和流式细胞术检测细胞死亡率和凋亡率的变化.结果 顺铂处理细胞0、3、6、12、24 h后LC3相对灰度值为0.33±0.04、1.13±0.18、1.92±0.25、0.89±0.12、0.58±0.08;在顺铂加入前、后1 h加入3-MA细胞的抑制率分别为(33.58±2.45)%、(21.37±1.21)%,细胞凋亡率分别为(18.30±0.92)%、(12.80±0.61)%.结论 顺铂可以诱导食管癌EC9706细胞自噬,在顺铂给药前期,自噬对细胞有保护作用,可以延迟凋亡的发生;而在顺铂给药后期,自噬则可能作为一种细胞死亡程序,进一步促进细胞死亡.

Role of autophagy in esophageal carcinoma cell line induced by cisplatin

Objective To explore the expression of autophagy in EC9706 cell line with treatment of Cisplatin, and study whether the expression level of autophagy is related to the death induced by Cisplatin.Methods EC9706 cells in logarithmic growth phase were treated with Cisplatin for 0, 3, 6, 12, 24 h,and then Western blotting was used to detect microtubule - associated protein 1 light chain 3 ( LC3 ) expression levels. After treatment of EC9706 cells with 3-Methyladenine (3-MA) and Cisplatin for 24 h (3-MA was added 1 h before and 1 h after Cisplatin respectively), the apoptosis rate and cell death rate were examined by flow cytometry and CCK-8 assays. Results The protein average values of LC3 were 0.33 ±0.04, 1.13 ±0.18, 1.92 ±0.25, 0.89 ±0.12, 0.58 ±0.08 at 0, 3, 6, 12, 24 h after treatment with cisplatin, respectively. The inhibitory rate after treatment of cells with Cisplatin and 3-MA (3-MA was added 1 h before and 1 h after Cisplatin respectively) was (33.58 ±2. 45 )% , (21.37 ± 1.21 )% , and the apoptotic rate was ( 18.30 ± 0.92) %, ( 12.80 ± 0.61 ) %, respectively. Conclusion Cisplatin can induce autophagy in EC9706. In the early, autophagy could delay apoptosis. However, in the end, autophagy may promote cell death.