Research On Inhibition Activity Of H-PNAS-4 Against Ovarian Cancer Cell Lines In Vitro

Objective To study whether transfection with apoptosis related newgene—h-PNAS-4 would show antitumer activity in A2780 /DDP(DDPresistant),A2780(DDP sensitive) and skov-3 cell lines of ovarian cancer,and to identify whether h-PNAS-4 combined with hyperthermia will showsynergistic antitumer activity to them。Methods Transfected A2780 /DDP,A2780 and skov-3 cells withh-PNAS-4 by liposome, through transfection with green fluorescentprotein(GFP) to identify transfection ratio; the MTT assay has beendeveloped for quantitative evaluation of the proliferation of A2780/DDP,A2780 after transfection of plasmid-based h-PNAS-4 48h later; Agarose gelelectrophoresis and Hoechst were used to show apoptosis-inducing activity24h later; treated these cells with hyperthermia(43℃for 2hours) to make surewhether h-PNAS-4 combined with hyperthermia will show synergisticantitumer activity by Weeb coefficient.Results1. h-PNAS-4 can be successfully transfected into cell lines of ovarian cancer by liposome-Lipofectamine 2000.2. h-PNAS-4 shows great proliferation inhibition to ovarian cancer celllines: the inhibition rate to A2780 /DDP, A2780 is 48.32%,60.48 %respectively, compared with the controls, P＜0.05.3. h-PNAS-4 induces apoptosis in ovarian cancer cell lines: Agarose gelelectrophoresis (DNA LADDER) demonstrated ladder-like pattern of DNAfragments; in morphological changes including HOECHST-stainedcondensed nuclei (intact of fragmented), nuclear fragmentation.4. When h-PNAS-4 combined with hyperthermia, they show additioneffect in cell line A2780/DDP, but no such effect in cell line A2780.Conclusion h-PNAS-4 can be successfully transfected into celt lines ofovarian cancer by liposome-Lipofectamine 2000; h-PNAS-4 exhibits greatproliferation inhibition, induce apoptosis in ovarian cancer cell lines; andwhen h-PNAS-4 combined with hyperthermia, they show addition effect incell line A2780/DDP but no such effect in cell line A2780.