Preliminary Research About The Effect Of 9-nitrocamptothecin Loaded Liposomes In Cycle And Apoptosis Of Duct Cancer Cells TFK-1

Objective To compare the influence in cell cycle and apoptosis of 9-nitrocamptothecin liposomes(9-NC-LP) and 9-nitrocamptothecin(9-NC) in bile duct cancer cell TFK-1 ,meanwhile evaluate the feasibility of liposomes as 9-NC nanocarrie.Methods TFK -1 was incubated with different concentrations of 9-NC and 9-NC-LP (0ng/ml,0.4ng/ml, 4ng/ml, 40ng/ml, 400ng/ml, 4000ng/ml) for 48h,72h ,24h; theninhibition rate was measured by MTT method. the cell TFK -1 was incubated with different concentrations of 9-NC and 9-NC-LP(0ng/ml ,40ng/ml,80ng/ml,160ng/ml,320ng/ml,640ng/ml) for 24h, 48h, 72h, using flow cytometric to detection cell cycle of distribution and the rate of cell apoptosis, and the western blotting to detection CDK2, Cyclin D1, Cyclin E, Cyclin A, P27, P53, Caspase-3,Caspase-8,Caspase-9,AIFM-1,Bax,Bcl-2 expression . Results After TFK-1 was incubated for 24h,48h ,the inhibition rate had no significant difference in 9-NC group and 9-NC -LP group ;but after 72h,the inhibition rate of 9-NC-LP group was significantly higher than 9 - NC group.After incubated for 24h ,cell in phase S of 9 -NC group was more obviously increased then 9 -NC -LP groups(p < 0.05) , but after 48h and 72h,cell in phase S of 9 -NC -LP group was more obviously increased then 9 -NC groups(p < 0.05),meanwhile after 72h, Annexin V + PI - cell count of 9-NC-LP group was significantly higher than 9 - NC group. After TFK -1 was incubated for 24h , the expression of cyclin A (F = 20.20, P = 0.02), P27 (F = 5.12, P = 0.05), P53 (F = 101.85, P < 0.01) caspase - 3 (F = 25.37, P < 0.01), caspase - 9 (F = 11.42, P = 0.01), Bax/Bcl - 2 (F = 39.94, P < 0.01) was significant difference among three groups. After TFK -1 was incubated for 72h, the expression of cyclin A (F = 310.66, P < 0.01), cyclin E (F = 8.26, P = 0.02), cyclin D1 (F = 396.03, P < 0.01), P27 (F = 7.50, P = 0.02), P53 (F = 944.62, P < 0.01) was significant difference among three groups. Conclusion 9 - NC - LP had strongly inhibited the growth and proliferation of TFK-1 and significantly raised Bax/Bcl-2,.After TFK-1 cell line was incubated with drug for 24h, 9-NC showed a more potent modulating effects on TFK-1 cell cycle and cell cycle related protein expression than 9-NC-LP. however, modulating effects show no difference between 9-NC and 9-NC-LP when TFK-1 cell line was incubated with drug for 72h. Which means 9-NC-LP have a good controlled release effect and incorporation with liposomes do not reduce antitumor effect of 9-NC. We believe 9-NC-LP is a promising antitumor formulation.