Effects Of Progesterone Receptor Ser294 Phosphorylation On Proliferation And Apoptosis Of Ovarian Cancer Cells

Objective: To investigate the effect of progesterone on the phosphorylation status of progesterone receptor Ser294 in ovarian cancer cells and the effects of progesterone receptor Ser294 phosphorylation on proliferation and apoptosis of ovarian cancer cells.Methods:?1?The ovarian cancer HO8910?PR overexpression?and SKOV3?PR low expression?cells were divided into four groups: R5020 group,R5020+U0126 group,U0126 group and control group.R5020 group was added medium without drug,cells were replaced with the complete medium of 10^-6mol/LR5020 at 6h,18 h,30h and 42 h.cells were replaced medium without drug at 12 h,24h and 36 h.R5020+U0126 group was added medium of 10^-5mol/LU0126,cells were replaced with complete medium of10^-6mol/LR5020 at 6h,18 h,30h,42 h,12h,24 h,36h.cell were replaced with complete medium of 10^-5mol/LU0126.U0126 group was added with complete medium of10^-5mol/LU0126.The control group was given a complete medium without drug.The cells were collected at 48 h,the relative expression of phosphorylated PRA Ser294 and PRB Ser294 was detected by Western-Blot.Since only phosphorylation of PRA Ser294 and PRB Ser294 occurs in each of the PR isoforms,we used the trend of phosphorylation of PRA Ser294 and PRBSer294 to indicate the trend of PRSer294 phosphorylation.?2?The experimental group was the same as above.The proliferation inhibition rate and apoptosis rate of HO8910 and SKOV3 cells were detected by CCK8 and Annexin V/PI double staining flow cytometry.Results:?1?The SKOV3 and HO8910 cell lines of U0126 group levels of PRB Ser294 phosphorylation relative expression were 0.006±0.002 and 0.007± 0.008,and the relative expression of PRA Ser294 phosphorylation was not detected,SKOV3 and HO8910 cell lines in U0126 group,The relative expression levels of PRB and Ser294 phosphorylation were 0.005±0.001 and 0.006±0.001;the relative expression of PRA Ser294 phosphorylation was not detected,In R5020 group,SKOV3 and HO8910 cell lines relative expression levels of PRB Ser294 phosphorylation were?0.72±0.028 and 0.68±0.045?;the relative expression of PRA and Ser294 phosphorylation was?0.18±0.027 and0.19±0.028?,In R5020+U0126 group SKOV3 and HO8910 cell lines PRB phosphorylation of Ser294 expression levels were?0.52 ±0.033,0.37±0.012?;HO8910 cell line PRA Ser294 phosphorylation expression levels were 0.14±0.016;SKOV3 cells phosphorylation of PRA Ser294 was not detected.The relative expression of PRA Ser294 and PRB Ser294 phosphorylation in group R5020 was higher than that in the control group,?P<0.05?The relative expression of PRA Ser294 and PRB Ser294 phosphorylation in R5020+U0126group was lower than that in R5020 group,?P<0.05?.?2?SKOV3 and HO8910 cell lines in U0126 group proliferation inhibition rate were?9.27%,28.22%?,SKOV3 and HO8910 cell of R5020 group proliferation inhibition rate were?40.36%,53.15%?,SKOV3 and HO8910 cell of R5020+U0126 group proliferation inhibition rate were?29.65%,38.66%?.The inhibition rate of proliferation of two strains in R5020 group was higher than that in group R5020+U0126,and the inhibition rate of proliferation of two strains in R5020+U0126 group was higher than that in U0126 group,?P<0.05?.?3?SKOV3 and HO8910 cell lines of control group apoptosis were?9.57%,9.79%?,SKOV3 and HO8910 cell lines of U0126 group apoptosis rates were?12.37%,12.62%?,SKOV3 and HO8910 cell lines of R5020 group apoptosis rate were?21.66%,27.82%?,and SKOV3 and HO8910 cell lines apoptosis of R5020+U0126 group were?14.11%,15.35%?.The apoptosis rate of SKOV3 and HO8910 cell lines of R5020 group was higher than control group and R5020+U0126 group?P<0.05?,the apoptosis rate of R5020+U0126 group was higher than that of U0126 group?P<0.05?.Conclusion: Progesterone can increase the relative expression of PRB Ser294 and PRA Ser294 phosphorylation in ovarian cancer HO8910 and SKOV3 cells.PRB Ser294 and PRA Ser294 phosphorylation increased,progesterone inhibited proliferation and apoptosis of ovarian cancer cell lines.