摘要
目的:探究Fas在胃癌细胞顺铂耐药中的作用及可能的作用机制。方法:Western blot及RT-q PCR检测胃癌细胞株SGC-7901及胃癌耐药细胞株SGC-7901/DDP中Fas的表达情况。构建Fas过表达腺病毒载体,上调胃癌耐药细胞株SGC-7901/DDP中Fas的表达,CCK-8检测细胞活力,流式细胞术检测细胞周期及细胞凋亡,Western blot法检测Fas、P38/p-P38、JNK/p-JNK、cleaved caspase-8/caspase-8和cleaved caspase-3/caspase-3蛋白水平。结果:耐药细胞株SGC7901/DDP中Fas的mRNA及蛋白表达水平均明显低于亲本细胞株;并且在亲本细胞株SGC7901中,Fas的蛋白表达水平随着顺铂的浓度增加不断降低。过表达Fas可抑制胃癌耐药细胞SGC7901/DDP的细胞活力,并诱导其凋亡;同时上调p-P38、p-JNK、cleaved caspase-8和cleaved caspase-3的蛋白水平。结论:胃癌顺铂耐药细胞中过表达Fas可恢复细胞对顺铂的敏感性,促进细胞凋亡并抑制细胞生长,其机制可能与JNK和P38MAPK信号通路的激活有关。
AIM： To study the effect of Fas on cisplatin resistance in stomach cancer cells and its possible mechanisms. METHODS： The expression of Fas at mRMA and protein levels in SGC-7901 cells and SGC-7901 / DDP cells was determined by RT-q PCR and Western blot. Fas-containing adenovirus vector was transfected into the SGC-7901 / DDP cells to upregulate Fas expression. The cell viability was detected by CCK-8 assay. The cell cycle and cell apoptosis were analyzed by flow cytometry. The protein levels of Fas,P38 / p-P38,JNK / p-JNK,cleaved caspase-8 / caspase-8 and cleaved caspase-3 / caspase-3 were detected by Western blot. RESULTS： The expression of Fas at both mRNA and protein levels was significantly downregulated in the SGC-7901 / DDP cells. Fas expression was decreased by cisplatin in a dose-dependent manner in the SGC-7901 cells. Overexpression of Fas suppressed the viability and induced apoptosis in the SGC-7901 / DDP cells,and upregulated the protein levels of p-P38,p-JNK,cleaved caspase-8 and cleaved caspase-3. CONCLUSION：Overexpression of Fas increases the sensitivity of the SGC-7901 / DDP cells to cisplatin,and inhibits the cell growth and promotes cell apoptosis. The mechanism may be related to the activation of JNK and P38 pathway.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2016年第9期1568-1573,共6页
Chinese Journal of Pathophysiology
基金
温州市科技计划项目(No.2012S0441
No.Y20130046
No.Y20140296)
关键词
FAS
胃癌
顺铂耐药
细胞凋亡
Fas
Stomach cancer
Cisplatin resistance
Cell apoptosis
