Effects Of Estrogen And Cisplatin On Growth Of Human Ovarian Cancer Cell Line HO-8910 In Vitro

ObjectiveTo investigat the effects of estrogen and cisplatin on growth of human ovarian cancer cell line HO-8910 in vitro.MethodsHuman ovarian cancer cell line HO-8910 were cultured in vitro. The treated groups consisted of estradiol group,cisplatin group and estradiol plus cisplatin group.The proliferational of HO-8910 cells was determined by methyl thiazolyl tetrazolium(MTT) method.Cell cycle and apoptotic percentage were detected by Flow CytometryMorphological and ultrastructure changes were observed by light and confocal laser microscop.Results1.Cell morphology Observation :(1) Observation by inverted microscope:Growth behavior of HO-8910 cells in control group and estrogen group was well. Extension cells showed like flagstone. As time went on, cell number increased graduallyand contact between the cells came to tightness. In E2 plus cisplatin group, the cell number was less than that of E2 group and control group but the cell form was acceptable when cells were incubated for 24 h. As time went on, vacuolecould be seen in cytoplasm. When incubated for 72 h, with cells shrinking, cell volume was smaller and intercellularspace enlarged. A few cells suspended in the culture medium.Some cells had broken nucleus but cytoplasm was still integrated.(2)Observastion by confocal laser microscop:A few apoptosis cells were observed in the control group.Comparing with control group,there was not significant difference in cisplitin group and E2 group . A lot of typical apoptosic cells were observed in E2 plus cisplitin group,the dyeing of cell membrane enhanced obviously,the intensity of fluorescence becamed more bright and different.2. Effects of 17-βE2 and17-βE2 plus cisplitin on the proliferation of human ovarian cancer cell line HO-8910:(1) Effects of 17-βE2 on the proliferation of human ovarian cancer cell line HO-8910: After 24 hours,the growth of HO-8910 cells was stimulated slightly by 17-βE2. But comparing with control group, there was no significant difference (P>0.05). After 48 hours,the growth of HO-8910 cell line was stimulated by 17-βE2 when its concentration was 10×10-9~50×10-9mol/L .The stimulative rate was 22.8% and 26.5%.Comparing with control group,there was significant difference (P<0.05) .After 72 hours, the role of 17-βE2 was stable. Comparing with control group, there was no significant difference (P>0.05).(2) Effects of 17-βE2 plus cispitin on the proliferation of human ovarian cancer cell line HO-8910: After 24 hours,the growth of HO-8910 cells was inhibited slightly by 17-βE2 plus cisplitin.The inhibitision was obviously when concentration of E2 was 50×10-9mol/L. Comparing with control group,there was significant difference (P<0.05). After 24 hours, the growth of HO-8910 cells was inhibited obviously by 17-βE2 plus cisplitin when concentration of E2 was 5×10-9mol/L ~50×10-9mol/L. Comparing with control group,there was significant difference (P<0.05) and extremely significant difference (P<0.01). .After 72 hours, the role of 17-βE2 plus cisplitin was stable. Comparing with control group, there was no significant difference (P>0.05).(3) The synergistic action of 17-βE2 plus cisplitin on the proliferation of human ovarian cancer cell line HO-8910:The effect showed additive when cisplitin combined with 1×10-9 17-βE2 and synergistic when cisplitin combined with5×10-9 ~50×10-917-βE23. Effects of 17-βE2 and17-βE2 plus cisplitin on cell cycle and apoptotic percentage of human ovarian cancer cell line HO-8910:(1) FCM showed that cell cycle distribution and apoptotic percentage were changed. After treatment with E2, the proportion of S phase was increased significantly, while the apoptotic percentage was reduced. The proportion of G0/G1 phase in cisplitin group was increased and the proportion of S phase was decreased, while the apoptotic percentage was increased.In 17-βE2 plus cisplitin group, the proportion of S phase was further decreased, while the apoptotic percentage was increased obviously.(2) Test of cell apoptosis by Annexin-V/PI staining method:①The apoptosic rate of HO-8910 cell treated by diffcrent concentration of E2 was 6.3%,6.0%,5.6%,5.1%. Comparing with control group, there was no significant difference (P>0.05). It is showed that E2 couldn′t induce the apoptosis of human ovarian cancer cell line HO-8910.②After treated by cisplitin, The apoptosic rate of HO-8910 cell was 15.6%,the difference was significant(P<0.05).③The apoptosic rate of HO-8910 cell treated by diffcrent concentration of E2 plus cisplitin was 16.8%,29.7%,31.2%,44.0%. Comparing with control group, there was significant difference (P<0.01).Conclusion1.17-βE2 couldn′t inhibit the apoptosis in human ovarian cancer cell line HO-8910 in vitro,but could enhance its proliferation.2. 17-βE2 plus cisplitin could change cell cycle distribution and apoptotic percentage.The proportion of S phase was further decreased, while the apoptotic percentage was increased obviously .It is suggested that 17-βE2 plus cisplitin could improve sensitivity and enhance the effect of chemotherapy.3.Treated with low dose estrogen to ovarian cancer patient after operation could improve quality of life.