Fas信号通路在维生素E琥珀酸酯诱导人胃癌SGC-7901细胞凋亡中的作用

Roles of Fas signaling pathway in vitamin E succinate-induced apoptosis in human gastric cancer SGC-7901 cells.

作者信息

Wu Kun, Li Yao, Zhao Yan, Shan Yu-Juan, Xia Wei, Yu Wei-Ping, Zhao Lan

机构信息

Department of Nutrition and Food Hygiene, Public Health School, Harbin Medical University, Heilongjiang Province, China.

出版信息

World J Gastroenterol. 2002 Dec;8(6):982-6. doi: 10.3748/wjg.v8.i6.982.

DOI:10.3748/wjg.v8.i6.982

PMID:12439910

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4656403/

Abstract

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.

摘要

目的

探讨Fas信号通路在维生素E琥珀酸酯诱导人胃癌SGC - 7901细胞凋亡中的作用。

方法

人胃癌SGC - 7901细胞分别用5、10、20mg·L⁻¹的维生素E琥珀酸酯（VES）处理，以琥珀酸和维生素E作为溶剂对照，仅以条件培养基作为未处理（UT）对照。通过DAPI染色观察凋亡形态。采用蛋白质印迹分析检测Fas、FADD和半胱天冬酶 - 8蛋白的表达。细胞分别用Fas和FADD反义寡核苷酸瞬时转染后，采用荧光法测定半胱天冬酶 - 8活性。

结果

DAPI染色显示VES处理后观察到细胞形态学凋亡变化。20mg·L⁻¹ VES处理24h和48h后，凋亡率分别为23.7%和89.6%。VES处理24h后，Fas、FADD和半胱天冬酶 - 8的蛋白水平明显呈剂量依赖性增加。用Fas反义寡核苷酸转染阻断Fas后，明显抑制了FADD蛋白的表达。SGC - 7901细胞用Fas和FADD反义寡核苷酸转染后，半胱天冬酶 - 8活性明显降低（P<0.01），而Fas阻断的作用大于FADD。