| Objective: To observe the effects of progesterone on the proliferation and metastasis of ovarian cancer SKOV3 and HO-8910 cells expressing different progesterone receptor membrane component 2 and explore their relationship with Wnt/?-catenin signaling pathway and epithelial-mesenchymal transition-related markers.Methods: 1.Ovarian cancer cell lines SKOV3(PGRMC2 low expression)and HO-8910(PGRMC2 high expression)were cultured in vitro.Two ovarian cancer cells were intervened with different concentrations of progesterone.Each cell was divided into 4groups: control group(0 ?mol/L),experimental groups(10 ?mol/L,20 ?mol/L,40 ?mol/L).After treated with progesterone on 24 h,48 h,72 h,the survival rate of SKOV3 and HO-8910 cells were detected by Cell Counting Kit-8;2.After treated with progesterone on48 h,the morphological changes of the two cells were observed by inverted microscope;3.After treated with progesterone on 48 h,the cell cycle distribution of the two cells were detected by Flow Cytometry;4.After treated with progesterone on 48 h,the cell migration and invasion ability of the two cells were detected by Transwell assays;5.After treated with progesterone on 48 h,the expressions of PGMRC2,?-catenin and E-cadherin of the two cells were detected by Western Blot.Results: 1.The results of CCK-8 showed that progesterone can inhibit the proliferation of ovarian cancer cells in a time-dose manner,and the effect on inhibiting proliferation of HO-8910 cell was more significant than that of SKOV3 cell(P<0.05);2.After 48 h of treatment with different concentrations of progesterone for the two ovarian cancer cells,the cell morphology gradually changed from paving stone-like morphology such as circular and polygonal to long fusiform with increased gap.With the increase of progesterone concentration,the number of adherent cells gradually decreased,suspended cells gradually increased,and the morphological changes of HO-8910 cell were more obvious than SKOV3 cell;3.Flow cytometry results showed that with the increase of progesterone concentration,the ratio of G0/G1 phase of the same ovarian cancer cell gradually increased(P<0.05),excluding the difference between the concentration of 10 ?mol/L and 20 ?mol/L,and the ratio of S phase gradually decreased(P<0.05),excluding the difference betweenthe concentration of 10 ?mol/L and 20 ?mol/L of SKOV3 cell.Moreover,the G0/G1 phase ratio of the HO-8910 cell control group and each experimental group was higher than that of SKOV3 cell(P<0.05);4.The results of Transwell migration and invasion experiments showed that the number of cells crossing the polycarbonate membrane in the control group of the same ovarian cancer cell was significantly higher than that in the experimental groups(P<0.05),and the metastasis ability of cells decreased with increasing progesterone concentration.With the same time and concentration of progesterone,the number of SKOV3 cell passing through the polycarbonate membrane was significantly higher than that of HO-8910(P<0.05);5.Western Blot results showed that compared with the control group,the levels of the PGRMC2 protein expression in the experimental groups in HO-8910 and SKOV3 cells were not statistically significant(P>0.05),but the PGMRC2 protein expression in HO-8910 cell was significantly higher than that in SKOV3 cell(P<0.05).Compared with the control group,the expression of ?-catenin protein in each experimental group of the two cell lines showed a decreasing trend(P<0.05),and the expression of E-cadherin protein showed an increasing trend(P<0.05).With the increase of progesterone concentration,the expression of ?-catenin decreased and the expression of E-cadherin increased(P<0.05),excluding the difference between the concentration of 10?mol/L and 20 ?mol/L,and With the same time and concentration of progesterone,the expression level of ?-catenin protein in SKOV3 cell was higher than that in HO-8910,and the expression level of E-cadherin protein was lower than that in HO-8910(P<0.05).Conclusion:1.The expression level of PGMRC2 protein is different in ovarian cancer cell lines SKOV3 and HO-8910 with different differentiation levels.Progesterone can inhibit the proliferation,migration and invasion of HO-8910 cell line with high PGRMC2 expression more obviously,compared with SKOV3 cell line with low PGRMC2 expression,suggesting that PGRMC2 may be a tumor suppressor;2.Progestogen may downregulate the activity of Wnt/?-catenin signaling pathway through PGRMC2,causing up-regulation of E-cadherin protein,and blocking the cell cycle progression of G0/G1 phase,thereby affecting the proliferation,migration and invasion of ovarian cancer SKOV3 and HO-8910 cells. |
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