细胞自噬在三氧化二砷诱导肝癌细胞凋亡中的作用研究

发布时间：2018-04-13 09:04

本文选题：肝细胞癌 + 三氧化二砷　；参考：《四川大学学报(医学版)》2017年02期

【摘要】：目的研究细胞自噬在三氧化二砷(ATO)诱导的肝癌细胞死亡中的作用,观察调控细胞自噬对ATO诱导的细胞凋亡有何影响。方法以人肝癌HepG_2细胞为研究对象,在ATO作用于HepG_2细胞的基础上,加入自噬激动剂〔雷帕霉素(Rap)〕和自噬抑制剂〔三甲基腺嘌呤(3-MA)〕进行调控。以MTT法检测细胞生存率,透射电镜观察自噬体结构,免疫印迹法分析自噬相关蛋白LC-3和Beclin1的表达;流式细胞术检测细胞凋亡。结果5~20μmol/L ATO与10 mmol/L 3-MA联用时HepG_2细胞存活率比同剂量ATO单独作用时降低(P0.05),凋亡率增加(P0.05),细胞内呈现典型凋亡反应特征,自噬蛋白LC3和Beclin1表达低于ATO单独作用组(P0.05)。相反,5~20μmol/L ATO与0.25μmol/L Rap联用时,HepG_2细胞的存活率比ATO单独作用时显著增加(P0.05),凋亡率降低(P0.05),细胞内出现大量自噬体结构,自噬蛋白LC3和Beclin1的表达较ATO单独作用组有增加趋势,但差异无统计学意义(P0.05)。结论特异性自噬抑制剂能显著增强ATO杀伤肝癌细胞的敏感性,其机制与增强肝癌细胞凋亡有关。
[Abstract]:Objective to investigate the role of autophagy in the apoptosis of hepatoma cells induced by arsenic trioxide (ATO) and to investigate the effect of autophagy on apoptosis induced by ATO.Methods Human hepatocellular carcinoma (HepG_2) cells were treated with ATO on the basis of HepG_2 cells. Autophagy agonists (rapamycin Raphe) and autophagy inhibitor (trimethyladenine 3-MAG) were added to control HepG_2 cells.The cell survival rate was detected by MTT method, the structure of autophagy was observed by transmission electron microscope, the expression of LC-3 and Beclin1 was detected by Western blotting, and apoptosis was detected by flow cytometry.On the contrary, the survival rate of HepG2 cells treated with 20 渭 mol/L ATO and 0.25 渭 mol/L Rap was significantly higher than that of ATO alone, the apoptotic rate decreased, and a large number of autophagy structures appeared. The expression of autophagy LC3 and Beclin1 increased compared with that of ATO alone.But the difference was not statistically significant (P 0.05).Conclusion the specific autophagy inhibitor can significantly enhance the sensitivity of ATO to the killing of hepatoma cells, and its mechanism is related to the enhancement of apoptosis of hepatoma cells.
【作者单位】：四川大学华西公共卫生学院环境卫生与职业医学系;
【基金】：国家自然科学基金面上项目(No.81372945) 四川省卫计委科研课题(No.150109)资助
【分类号】：R735.7

【参考文献】