摘要
目的探讨维生素E琥珀酸酯(vitamin E succinate,VES)对人胃癌SGC-7901细胞线粒体凋亡通路中第二种线粒体来源半胱氨酸蛋白酶激活剂(secondmitochondria-derivedactivatorofcaspases,Smac)、X连锁凋亡抑制蛋白(X-linked inhibitor of apoptosis protein,XIAP)基因和蛋白表达的影响及其诱导凋亡机制。方法不同浓度VES(5、10、15、20μg/ml)处理人胃癌细胞24h,0.01%的无水乙醇设为对照组。CCK-8检测细胞增殖活性;实时荧光定量PCR检测线粒体相关凋亡分子Smac、XIAP mRNA表达变化;Western blot检测线粒体相关凋亡分子Smac、XIAP蛋白表达水平的差异;Annexin-V/PI双染流式细胞术检测各组细胞凋亡率。结果与对照组相比,不同浓度VES(5、10、15、20μg/ml)作用胃癌细胞后的细胞增殖率分别为94.67%±0.33%、85.28%±0.82%、67.63%±1.3%、52.11%±2.3%,VES抑制细胞增殖的效果随VES浓度升高而增强(P<0.05);Smac mRNA随着VES剂量的增加而增加(P<0.05),在15μg/ml达到峰值(P<0.05),XIAPmRNA随着VES剂量的增加而下降(P<0.05);XIAP蛋白在5、10μg/mlVES处理组表达量差异无统计学意义,在15、20μg/ml VES处理组,表达量差异有统计学意义(P<0.05),蛋白表达量随着浓度升高(P<0.05),Smac蛋白蛋白表达量随着VES剂量的增加明显升高(P<0.05);不同浓度VES(5、10、15、20μg/ml)处理组凋亡率依次为2.06%±0.21%、8.17%±0.56%、13.30%±0.33%、19.82%±0.95%、34.45%±1.78%,VES诱导细胞凋亡的效果随VES作用浓度升高而升高(P<0.05)。结论VES可以抑制胃癌细胞增殖、诱导细胞凋亡,在转录和翻译水平上上调Smac的表达、抑制XIAP的表达。
Objective To investigate the effects of vitamin E succinate(vitamin E succinate,VES),on the second mitochondria-derived activator of caspases(Smac)in the mitochondrial apoptosis pathway and X-linked inhibitor of apoptosis protein(XIAP)gene and protein expression in human gastric cancer SGC-7901 cells and their apoptosis-inducing mechanism.Methods Human gastric cancer SGC-7901 cells were treated with VES(5,10,15,20μg/ml)at different concentrations for 24 h,and 0.01%anhydrous ethanol was set as the control group.CCK-8 kit was used to detect the proliferation activity of cells.The mRNA expression of mitochondria-related apoptotic molecules Smac and XIAP was detected by real-time fluorescence quantitative PCR.Western blot was used to detect the protein expression of Smac and XIAP.Annexin-v/PI double staining flow cytometry was used to detect the apoptosis rate.Results Compared with the control group,the proliferation rates of gastric cancer cells treated by VES(5,10,15,20μg/ml)were 94.67%±0.33%,85.28%±0.82%,67.63%±1.3%,52.11%±2.3%,respectively.The inhibitory effect of VES on cell proliferation was VES concentration dependent(P<0.05).Smac mRNA expression increased after VES treatment(P<0.05),and reached the peak value at VES 15μg/ml(P<0.05).XIAP mRNA expression decreased with the increase of VES dose(P<0.05).There was no significant difference in the expression level of XIAP protein between 5μg/ml and 10μg/ml VES treatment groups.The expression levels in 15μg/ml and 20μg/ml VES treatment groups were significantly different(P<0.05).The Smac protein expression level increased in a VES concentration dependent manner.The apoptosis rates of VES(5,10,15,20μg/ml)treatment groups were 2.06%±0.21%,8.17%±0.56%,13.30%±0.33%,19.82%±0.95%,34.45%±1.78%,respectively.The apoptosis-inducing effect of VES was significantly depended on VES concenteation(P<0.05).Conclusion VES can inhibit the proliferation and induce apoptosis in gastric cancer cells,by up-regulating the expression of Smac and inhibiting the expression of XIAP at the transcription and translation levels.
作者
冯万芳
张馨竹
高杨叶
王弈丹
侯丽颖
FENG Wan-fang;ZHANG Xin-zhu;GAO Yang-ye;WANG Yi-dan;HOU Li-ying(School of Public Health,North China University of Science and Technology,Tangshan 063210;School of Science,North China University of Science and Technology,Tangshan 063210,China)
出处
《营养学报》
CAS
CSCD
北大核心
2021年第5期450-455,共6页
Acta Nutrimenta Sinica
基金
河北省自然科学基金(No.H2019209453)
华北理工大学博士科研项目(No.BS2017058)
华北理工大学大学生创新创业训练计划(No.X2019107)
作者简介
冯万芳(1998-),女,医学学士,E-mail:526673586@qq.com;通讯作者:侯丽颖,E-mail:houliying1987@163.com
