Roles of Fas signaling pathway in vitamin E succinate-induced apoptosis in human gastric cancer SGC-7901 cells.
Roles of Fas signaling pathway in vitamin E succinate-induced apoptosis in human gastric cancer SGC-7901 cells.
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DOI：
10.3748/wjg.v8.i6.982
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发表时间：
2002-12
影响因子：
4.3
通讯作者：
Kun Wu;Yao Li;Yan Zhao;Y. Shan;W. Xia;Wei-Ping Yu;Lan Zhao
中科院分区：
文献类型：
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作者：
Kun Wu;Yao Li;Yan Zhao;Y. Shan;W. Xia;Wei-Ping Yu;Lan Zhao
目的探讨Fas信号通路在维生素E琥珀酸诱导人胃癌SGC-7901细胞凋亡中的作用。方法用5、10、20 mg x L(-1)的VES处理人胃癌SGC-7901细胞，琥珀酸和维生素E作为载体对照，条件培养基仅作为未处理(UT)对照。通过DAPI染色观察细胞凋亡形态。应用蛋白质印迹分析来测量Fas、FADD和caspase-8蛋白的表达。分别用Fas和FADD反义寡核苷酸瞬时转染细胞后，通过荧光法测定caspase-8活性。结果DAPI染色观察VES处理后细胞形态学的变化。 20 mg·L(-1) VES处理24 h和48 h后，细胞凋亡率分别为23.7%和89.6%。 VES处理24小时后，Fas、FADD和caspase-8的蛋白水平明显增加，并呈剂量依赖性。 Fas反义寡核苷酸转染对Fas的阻断明显抑制了FADD蛋白的表达。 Fas和FADD反义寡核苷酸转染SGC-7901细胞后，caspase-8活性明显降低（P<0.01），而Fas的阻断程度高于FADD。结论 VES诱导人胃癌SGC-7901细胞凋亡涉及Fas信号通路，包括Fas、FADD和caspase-8的相互作用。
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 x 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 x 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.