The Research Of Targeting Therapy Of OSTP And Cisplatin Copolymer Agent In Ovarian Cancer A2780 Cells

Objective:In the previous research,we have screened the ovarian cancer specific targeting peptide OSTP by using phage display teconology in vivo,and have already verified the property.In this research,we had associated OSTP with DDP,and we had to study the targeting inhibition of this coupled object.Lay a foundation of the exploration of the new tumor molecular medicine.Methods:The company of Sangon Biotech in Shanghai had synthesised OSTP-DDP in chemical.Culture the ovarian cancer A2780 cells in vitro.Using cell counting Kit-8 to detected the growth inhibition effect of OSTP-DDP and DDP respectly,and counting for its IC50. Using tablet clone forming experiments to detected the growth inhibition effect of OSTP-DDP and DDP respectly.Using flow cytometry to detected the cycle and apoptosis effect of OSTP-DDP and DDP respectly. Using high performance liquid chromatography to detected the does that enter into the ovarian cancer A2780 cells so that we can directly detect the targeting of OSTP-DDP.Builded a nude-mouse tansplanted model of ovarian cancer A2780 cells(control group、DMSO group、DDP group、OSTP-DDPgroup) to detect the targeting of OSTP-DDP in vivo.We used blood routine examination and routine biochemistry to detect whether OSTP-DDP has a less toxicity in vivo.Results:1.we could know from mass spectrometry and high performance liquid chromatography that OSTP-DDP has been conjugated successfully.2. Cell counting Kit-8 indicated that using different dose of OSTP-DDP and DDP(10、20、40、80、160、320 μmol?L-1) to dealed with the ovarian cancer A2780 cells of 24h、48h、72h,both of them can inhibite the growth of the cells,and it depend on time and concertration.The effect of OSTP-DDP is better than DDP’s(p<0.05).It hint us OSTP-DDP has the effect of targeting inhibition of the ovarian cancer A2780 cells.3. Plated clone forming assay indicated that both OSTP-DDP and DDP can inhibited the growth of the cells,but there had no statistically significant differences between them(p>0.05).4.Using FCM we can know that after treat with OSTP-DDP and DDP,the cycle of the cells are all retardanted in G1 phase,after treat with72 h,the effect of OSTP-DDP is better than DDP’s(p<0.05).After treated by 24 h,48h,72 h, the apoptosis rates of OSTP-DDP group respectly are 7.51%、44.46%、52.73%,the apoptosis rates of DDP group respectly are 4.44%、13.66%、15.67%。The effect of cell apoptosis of OSTP-DDP is better than DDP’s(p<0.05).It remind us that OSTP-DDP has a sstrongtargeting effect to induce cell apoptosis.5.Using high performance liquid chromatography we know that after treat by 12 h,the dose of OSTP-DDP enter into cells are more than DDP’s.It is directly indicated that OSTP-DDP has targeting of ovarian cancer A2780 cells.6.A nude-mouse tansplanted model has established successfully,the experiment in vivo indicated that both of OSTP-DDP and DDP can inhibite the tumor,the effect of OSTP-DDP is better than DDP’s(p<0.05). Blood routine examination and routine biochemistry can decrease the toxicity(p<0.05).Conclusion: OSTP-DDP has a good targeting combination with ovarian cancer A2780 cells both in vivo and in vitro,and it can inhibited the growth and activity of tumor.It is hopefully to build a new method to targeting ovarian cancer,and can provide a new train of thought and helpful exploration to target ovarian cancer and other human cancers.