Effects Of AGO14699Alone And Combined With Cisplatin On Proliferation Of Human Ovarian Cancer Cell Line SKOV3and Cellular Apoptosis

Objective:To investigate the effect of AG014699combined with cisplatin on the proliferation,cell cycle and apoptosis of human ovarian cancer cell line SKOV3. So as to provide empirical and theoretical evidence of applying AGO14699as the combination with chemotherapy to treat ovarian cancer.Methods:Human ovarian cancer cell line SKOV3cultured in vitro were treated with different concentration of AGO14699,CDDP,AG014699combined with CDDP.Inverted microscope was used to detect apoptosis.The growth inhibition effect was investigated by MTT colorimetry, the distribution changes of cell cycles and the influences of cell apoptosis ratio were analyzed by the flow cytometry.Results:1.Under the microscope,the black control group cells grow even stick wall,good condition,even the size of the cells,the film clear cytoplasm and full.nucleoli and nuclear menbrane,outline obvious,and the adjacent cell growth confluent; Single drug AGO14699group see pseudopods retraction cells, cells go round, suspension, refraction sex difference, along with the drug concentration increases and role of the extension of time, the apoptotic cells increase gradually. Single drug CDDP group visible cells bad form, and with the increase of concentration and the extension of time, the limited growth is obvious; Along with the higher concentration of combined treatment group, not only cell suspension and refraction sex differences, the apoptotic cells increases gradually, but also the cell wall by bottle fall off, cells suspended connection piece.2.MTT showed that the different concentrations of AGO14699alone and combined with cisplatin exerted inhibitory effect on proliferation of SKOV3cells,and its effect presented a dose-dependent way.With the increasing concentration of AGO14699treatment,the inhibitory effects were enhanced,but with the extending time of treatment, the inhibitory effects were reduced.The differences had statistical significance(P<0.05);Different concentration of AG014699,10μmol/L、20μmol/L、50μmol/L、100μmol/L,acted respectively on SKOV3cell for24h、48h、72h,their spectrophotometries were0.398±0.038、0.348、0.038、0.291±0.0.046、0.235±0.033,0.39810.038、0.348±0.038、0.291±0.0.046、0.235±0.033,0.59210.041、0.545±0.043、0.48710.039、0.41810.028,0.74010.013、0.693±0.010、0.57510.010、0.521±0.010。Different concentration of AG014699, lOμmol/L、50μmol/L、100umol/L acted together with different concertration of CDDP,2.5、5、10μg/ml,on SKOV3cell for24hours,their spectrophotometries are0.523±0.013、0.490±0.010、0.417±0.025、0.344±0.011;0.500±0.019、0.386±0.020、0.356±0.023、0.303±0.008;0.327±0.015、0.294±0.007、0.257±0.022、0.218±0.020;0.222±0.013、0.214±0.018、0.199±0.090、0.164±0.018The concentration of50μmol/L of AG014699respectively with the concentration of2.5、5、10μg/ml CDDP acted together on SKOV3cell for24h, SKOV3cells inhibition rate from the original respectively3.92%,18.31%,34.57%rose to40.42%,49.69%, and58.23%. The q values of the combine medication were1.02,1.16and1.07.3.Flow cytometry showed that AG014699could block cell cycle in G0/G1phase. Combined with CDDP, AG014699could reinforce its effect and block cell cycle in G0/G1phase significantly.After treated with CDDP(2.5μg/ml)for24h,the apoptosis ratio of SKOV3cell was21.20±0.40.The effect changed obviously when AG014699(50μmol/L) combined with CDDP,the result raised up to30.58±0.50. The differences had statistical significance(P<0.05).Conclusion:AG014699can inhibit proliferation of SKOV3in vitro,block cell cycle,and induce cell apoptosis.Combined with CDDP, AGO14699can reinforce its effects.