摘要
AIM: To investigate the roles of Fas signaling pathway in vitamin E succinate-induced apoptosis in human gastric cancer SGC-7901 cells.METHODS: Human gastric cancer SGC-7901 cells were treated with VES at 5, 10, 20 mg@L-1, succinic acid and vitamin E as vehicle control and condition media only as untreated (UT) control. Apoptotic morphology was observed by DAPI staining. Western blot analysis was applied to measure the expression of Fas, FADD and caspase-8 proteins. After the cells were transiently transfected with Fas and FADD antisense oligonucleotides, respectively, caspase-8 activity was determined by flurometric method.RESULTS: The morphologically apoptotic changes were observed after VES treatment by DAPI staining. 23.7 % and 89.6 % apoptosis occurred after 24 h and 48 h of 20 mg@L-1 VES treatment, respectively. The protein levels of Fas, FADD and caspase-8 were evidently increased in a dose-dependent manner after 24 h of VES treatment. The blockage of Fas by transfection with Fas antisense oligonucleotides obviously inhibited the expression of FADD protein. After SGC-7901 cells were transfected with Fas and FADD antisense oligonucleotides, caspase-8 activity was obviously decreased (P＜0.01), whereas Fas blocked more than FADD.CONCLUSION: VES-induced apoptosis in human gastric cancer SGC-7901 cells involves Fas signaling pathway including the interaction of Fas, FADD and caspase-8.
AIM:To investigate the roles of Fas signaling pathway in vitamin E succinate-induced apoptosis in human gastric cancer SGC-7901 cells. METHODS:Human gastric cancer SGC-7901 cells were treated with VES at 5,10,20 mg·L^(-1),succinic acid and vitamin E as vehicle control and condition media only as untreated (UT)control.Apoptotic morphology was observed by DAPI staining.Western blot analysis was applied to measure the expression of Fas,FADD and caspase-8 proteins.After the cells were transiently transfected with Fas and FADD antisense oligonucleotides,respectively,caspase-8 activity was determined by flurometric method. RESULTS:The morphologically apoptotic changes were observed alter VES treatment by DAPI staining.23.7 % and 89.6 % apoptosis occurred alter 24 h and 48 h of 20 mg·L^(-1) VES treatment,respectively.The protein levels of Fas,FADD and caspase-8 were evidently increased in a dose-dependent manner alter 24 h of VES treatment.The blockage of Fas by transfection with Fas antisense oligonucleotides obviously inhibited the expression of FADD protein.After SGC-7901 cells were transfected with Fas and FADD antisense oligonucleotides,caspase-8 activity was obviously decreased (P<0.01),whereas Fas blocked more than FADD. CONCLUSION:VES-induced apoptosis in human gastric cancer SGC-7901 cells involves Fas signaling pathway including the interaction of Fas,FADD and caspase-8.
作者
Kun Wu Yao Li Yan Zhao Yu-Juan Shah Wei Xia Lan Zhao,Department of Nutrition and Food Hygiene,Public Health School,Harbin Medical University,Harbin 150001,Heilongjiang Province,China Wei-Ping Yu,Genetics Institute,Texas University of USA,Austin,USA
基金
National Natural Science Foundation of China,No.39970647
