维生素E琥珀酸酯诱导胃癌细胞自噬对细胞抗氧化功能的影响

EFFECT OF AUTOPHAGY INDUCED BY VITAMIN E SUCCINATE ON ANTIOXIDANT CAPACITY IN GASTRIC CANCER CELLS

目的探讨维生素E琥珀酸酯(VES)处理人胃癌SGC-7901细胞过程中自噬对细胞抗氧化功能的影响。方法不同剂量(0、5、10、15、20μg/ml)VES处理人胃癌SGC-7901细胞24h,Western blot检测自噬标志蛋白LC3和Beclin-1的表达情况,Real-time PCR检测细胞内抗氧化酶SOD、CAT和GPX mRNA的表达情况,ELISA检测GSH水平;RNA瞬时干扰阻断自噬关键基因BECN-1基因的表达后,以20μg/ml VES处理人胃癌SGC-7901细胞24h,Real-time PCR检测抗氧化酶SOD、CAT和GPX mRNA的表达情况,ELISA检测GSH水平。结果与对照组相比,随着VES作用剂量的增加,Beclin-1蛋白表达逐渐增高,可观察到LC3-Ⅰ向LC3-Ⅱ转化增加;Real-time PCR检测显示SOD、CAT和GPX等抗氧化酶的mRNA表达减少,ELISA显示细胞内GSH水平降低。阻断自噬关键基因BECN-1基因后,SOD、CAT和GPX等抗氧化酶的mRNA表达进一步减少(P<0.05),ELISA显示GSH水平进一步降低(P<0.05)。结论VES可诱导人胃癌SGC-7901细胞发生自噬并降低细胞内抗氧化物质的水平,抑制自噬后抗氧化酶的表达水平进一步降低,提示自噬可缓解VES引起的细胞氧化应激。

Objective To investigate the effect of autophagy on antioxidant capacity in human gastric cancer cells treated with vitamin E succinate(VES).Methods After the BECN-1 gene expression was blocked by RNA transient interference,human gastric cancer SGC-7901 cells were treated with VES at different doses(0,5,10,15,20 g/ml)for 24 hours.The expressions of autophagy marker proteins LC3 and Beclin-1 were detected by Real-Time PCR,and the expressions of antioxidant enzyme SOD,CAT and GPX mRNA were detected by real-time PCR,and GSH level by ELISA.Results Compared with control group,Beclin-1 protein expression increased in a VES dose dependent manner.An increase in the transformation from LC3-Ⅰto LC3-Ⅱwere also observed.Real-time PCR showed that the mRNA expression of antioxidant enzymes such as SOD,CAT and GPX decreased,and the level of GSH also decreased by ELISA.After blocking the key autophagy gene BECN-1,the mRNA expression of antioxidant enzymes such as SOD,CAT and GPX also was further reduced(P<0.05),and GSH level was further declined(P<0.05).Conclusion VES can induce autophagy in human gastric cancer SGC-7901 cells and reduce the level of intracellular antioxidants,and further reduce the level of antioxidants after the autophagy was inhibited,suggesting that autophagy can alleviate cellular oxidative stress induced by VES.[ACTA NUTRIMENTA SINICA,2021,43(6):550-555]