GOLPH3沉默对顺铂诱导的人上皮性卵巢癌 A2780／DDP 细胞凋亡的影响

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 －mTOR 蛋白表达的影响。Western blot 检测 mTOR 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 －mTOR 蛋白表达下调；LY294002和 Ra-pamycin 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／mTOR 激活促进顺铂诱导的 A2780／DDP 细胞凋亡。

Effect of GOLPH3 silencing on cisplatin -induced cell apoptosis in human epithelial ovar-ian cancer A2780/DDP cells

Objective:To explore the effect of Golgi phosphoprotein 3(GOLPH3)on cisplatin -induced cell apop-tosis in human epithelial ovarian cancer A2780 /DDP cells.Methods:After cisplatin treatment of A2780 and A2780 /DDP cells for 72h,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 48h,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 -mTOR proteins was determined by Western blot.The mTOR inhibitor Rapamycin and PI3K/Akt in-hibitor LY294002 were introduced to determine Caspase -3 expression by Western blot.Results:After cisplatin treat-ment for 72h,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 -mTOR proteins(P <0.05).Moreover,LY294002 and Rapamycin treatment promoted Caspase -3 expression,respectively(P <0.05).Conclusion:GOLPH3 silencing promoted cisplatin -in-duced cell apoptosis in human epithelial ovarian cancer A2780 /DDP cells through inhibiting Akt/mTOR activation.