Nedaplatin Induced Apoptosis In Human Ovarian Cancer Cisplatin-resistant Cell Lines In Vitro And Investigated Its Mechanism

Objective:To study the inhibition of nedaplatin(NDP) on human ovarian cancer cell lines SKOV3with intrinsic cisplatin-resistance and SKOV3/DDP with acquired cisplatin-resistance, to investigate its mechanism of inducing apoptosis and anticancer mechanism of nedaplatin,moreover, to provide experimental reference for clinical therapy of cisplatin-resistant ovarian cancerMethods:MTT assay was used to evaluate the inhibition in different concentration and time of nedaplatin treatment on SKOV3and SKOV3/DDP cells,then calculated median inhibitory concentration(IC50). Cells apoptosis and cells cycle of cells before and after treatment of drug were detected by flow cytometry(FCM).The changes of Bc1-2、Bax、Caspase3、Caspase9. MMP-2in mRNA levels were quantified by RT-PCR. Inverted phase contrast microscope and fluorescent staining were applied to observate the morphological changes of ovarian cancer cells. The statistical analysis was executed by SPSS16.0software, using variance analysis and t-test to analysis data, statistically significant level was "a=0.05".Results:1、Nedaplatin inhibited the growth of SKOV3and SKOV3/DDP cells significantly in time-dose-dependent fashion(P<0.05).2、FCM showd that apoptotic rate and the percentage of S-phase cells were tend to increase with time and concentration of nedaplatin (P<0.05), G0/G1-phase cells were tend to decrease (P<0.05), and G2/M-phase cells were no change (P>0.05)3、With the inverted phase contrast microscope and fluorescent staining, we observed the morphological changes of apoptotic SKOV3and SKOV3/DDP cells after treatment of nedaplatin..4、Compared with control groups,there were a decrease in Bcl-2and MMP-2mRNA expression,but an increase in Bax、Caspase3and Caspase9(P<0.05).Conclusion:1、NDP can inhibit proliferation of SKOV3and SKOV3/DDP cells in vitro significantly.The effect may be ascribed to inducing cells apoptosis, the S-phase arrest and the down-regulation of Bcl-2expression,up-regulation of Bax、 Caspase3and Caspase9expression.2、NDP cuts MMP-2expression,so it is advantageous to reduce ovarian cancer cells’invasion force.3、NDP can inhibit human ovarian cancer cisplatin-resistant cells, in addition,it is profit to reduce ovarian cancer cells’invasion force,so NDP is expected to become effctive platinum drug for therapy of ovarian cancer with cisplatin-resistance clinically.