摘要
目的:分析索拉菲尼对肝细胞癌凋亡及自噬相关蛋白表达的影响,及肝细胞癌对其耐药的机制。方法:常规培养人肝癌SMMC-7721细胞,以1.25、2.5、5、10、20μmol/L索拉菲尼处理细胞,观察其对细胞增殖的影响。观察索拉菲尼和自噬抑制剂3-甲基腺嘌呤(3-MA)对细胞凋亡和自噬,及其相关蛋白表达的影响。结果:随着索拉菲尼浓度增加,SMMC-7721细胞存活率逐渐降低,且呈时间依赖性(P<0.05),10μmol/L索拉菲尼对SMMC-7721增殖抑制作用相对较强,因此选用10μmol/L索拉菲尼进行后续研究;与空白对照组比较,索拉菲尼组细胞平均凋亡率显著增加,Bcl-2蛋白表达水平显著降低,Bax蛋白表达水平显著升高,细胞内自噬体显著增加,LC3-Ⅰ、LC3-Ⅱ和Atg5蛋白表达水平显著升高,差异均有统计学意义(P<0.05);加入3-MA后,SMMC-7721细胞和经索拉菲尼处理的SMMC-7721细胞平均凋亡率显著增加,细胞内自噬体显著减少,Bcl-2、LC3-Ⅰ、LC3-Ⅱ和Atg5蛋白表达水平较加入3-MA前显著降低,Bax蛋白表达水平显著升高,差异均有统计学意义(P<0.05)。结论:索拉菲尼可通过上调促凋亡蛋白和自噬相关蛋白水平,促进细胞凋亡和诱导细胞自噬,抑制自噬可增强索拉菲尼对肝癌细胞的杀伤效应,降低肝细胞癌对索拉菲尼的耐药。
Objective: To investigate the effect of sorafenib on apoptosis and autophagy related protein expression in hepatocellular carcinoma, and analyze its mechanism of drug resistance.Methods: Human hepatoma cell SMMC-7721 cells were cultured routinely, treated with different concentrations of sorafenib(1.25μmol/L, 2.5μmol/L, 5μmol/L, 10μmol/L, 20 μmol/L). Cell proliferation was observed. Then the effects of sorafenib and autophagy inhibitor 3-methyladenine(3-MA)on apoptosis, autophagy and the expression of related proteins were also observed. Results: With the increase of sorafenib concentration, the survival rate of SMMC-7721 cells decreased gradually in a time-dependent manner. 10 mol/L sorafenib was used in the follow study for its relatively significant inhibition effect on SMMC-7721 proliferation. Compared with the blank control group, the average apoptosis rate of cells in the Sorafenib group was significantly increased, the expression level of Bcl-2 protein was significantly decreased, and the expression level of Bax protein was significantly increased, autophagy increased significantly, and LC3-I, LC3-II and Atg5 protein expression levels were significantly increased(P<0.05);After adding 3-MA, the average apoptosis rate of SMMC-7721 cells and SMMC-7721 cells treated with sorafenib increased significantly, the autophagy decreased significantly, the protein expression levels of Bcl-2, LC3-I, LC3-II and Atg5 were significantly decreased, and the expression level of Bax protein was significantly increased(P<0.05). Conclusion:Sorafenib can promote apoptosis and induce autophagy by up regulating the levels of pro-apoptotic protein and autophagy related protein. Inhibition of autophagy can enhance the apoptosis of sorafenib on hepatoma cells and reduce the resistance of hepatocellular carcinoma to sorafenib.
作者
李晓明
姚立鹏
程丽慧
LI Xiao-ming;YAO Li-peng;CHENG Li-hui(Department of Hepatobiliary Surgery,Sanya People's Hospital,Sanya 572000,China;Department of Urology,Sanya People's Hospital,Sanya 572000,China)
出处
《中国现代普通外科进展》
CAS
2022年第1期7-11,共5页
Chinese Journal of Current Advances in General Surgery
关键词
肝细胞癌
索拉菲尼
凋亡
自噬
耐药性
Hepatocellular carcinoma
Sorafenib
Apoptosis
Autophagy
Drug resistance
