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.
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