摘要
目的:探究GOLPH3对顺铂诱导的人上皮性卵巢癌A2780/DDP细胞凋亡的影响。方法:不同浓度顺铂处理A2780和A2780/DDP细胞72h,MTT检测半数抑制浓度IC50,Western blot检测GOLPH3的表达。将培养细胞分为4组:A2780组及A2780/DDP组(对照组、Scrambled siRNA组、GOLPH3 siRNA组)。40μmol/L顺铂处理48h后,MTT检测对细胞活性的影响,流式细胞术检测对细胞凋亡的影响,Western blot检测对Caspase-3、p-Akt和p-m TOR蛋白表达的影响。Western blot检测m TOR抑制剂雷帕霉素(Rapamycin)和PI3K/Akt抑制剂(LY294002)对Caspase-3蛋白表达的影响。结果:顺铂处理72h时,A2780和A2780/DDP细胞的IC50分别为9.8μmol/L和60.14μmol/L。与A2780组相比,A2780/DDP组GOLPH3蛋白表达明显增加(P<0.05)。GOLPH3 siRNA转染可显著下调A2780/DDP细胞中GOLPH3蛋白的表达(P<0.05)。顺铂处理后,与A2780/DDP对照组相比,A2780组和A2780/DDP细胞GOLPH3 siRNA转染组细胞活性显著降低,顺铂敏感性增加,细胞凋亡率和Caspase-3蛋白表达上调,p-Akt和p-m TOR蛋白表达下调;LY294002和Rapamycin处理均显著增加Caspase-3蛋白表达(P<0.05)。结论:GOLPH3沉默可通过抑制Akt/m TOR激活促进顺铂诱导的A2780/DDP细胞凋亡。
Objective：To explore the effect of Golgi phosphoprotein 3（ GOLPH3） on cisplatin-induced cell apoptosis in human epithelial ovarian cancer A2780 / DDP cells.Methods：After cisplatin treatment of A2780 and A2780 /DDP cells for 72 h,the half inhibitory concentration（ IC50） was detected by MTT assay,and the GOLPH3 expression was detected by Western blot.Cultured cells were divided into four groups：A2780 group,A2780 / DDP group（ control group,Scrambled siRNA group,GOLPH3 siRNA group）.After 40μmol / L cisplatin treatment for 48 h,the cell viability was detected by MTT assay,the cell apoptosis was determined by flow cytometry,and the expression of Caspase-3,p-Akt,and p-m TOR proteins was determined by Western blot.The m TOR inhibitor Rapamycin and PI3 K / Akt inhibitor LY294002 were introduced to determine Caspase-3 expression by Western blot.Results：After cisplatin treatment for 72 h,the IC50 of cisplatin in A2780 and A2780 / DDP group was 9.8μmol / L and 60.14μmol / L,respectively.Compared to A2780 group,the GOLPH3 expression in A2780 / DDP group was significantly increased（ P〈0.05）.GOLPH3 siRNA transfection reduced GOLPH3 expression in A2780 / DDP cells（ P〈0.05）.After cisplatin treatment,compared to A2780 / DDP control group,both A2780 group and A2780 / DDP GOLPH3 siRNA group exhibited lower cell viability and elevated cisplatin sensitivity,up-regulated cell apoptosis rate and Caspase-3 expression,and down-regulated expression of p-Akt and p-m TOR proteins（ P〈0.05）.Moreover,LY294002 and Rapamycin treatment promoted Caspase-3 expression,respectively（ P〈0.05）.Conclusion：GOLPH3 silencing promoted cisplatin-induced cell apoptosis in human epithelial ovarian cancer A2780 / DDP cells through inhibiting Akt / m TOR activation.
出处
《现代肿瘤医学》
CAS
2016年第17期2700-2704,共5页
Journal of Modern Oncology
关键词
上皮性卵巢癌
高尔基体磷蛋白3
顺铂
细胞凋亡
epithelial ovarian cancer
Golgi phosphoprotein 3
cisplatin
cell apoptosis
