The Effects Of MPA And MG132 On Proliferation And Apoptosis Of Ovarian Cancer Cells With Different PR Expression

Objective: With the wide application of hormone therapy in the treatment of breast cancer and endometrial cancer,progesterone and progesterone receptors have become an important research direction in the treatment of ovarian cancer.MG132 is an inhibitor of proteasome pathway,and it has been shown that it is a potential anticancer drug.The purpose of this study was to investigate the effects of MPA and MG132 on the proliferation and apoptosis of ovarian cancer cells with different expression of PR to provide a new experimental basis for the clinical diagnosis and treatment of ovarian cancer.Methods: Choice the HO-8910(PR high expression)and SKOV3(PR low expression)cells as the research objects.(1)Different concentration of MPA(0,1,20,50,100 ?mol/L),MG132(0,1,2.5,5,10 ?mol/L)were used to treat alone for 24-72 h.Inhibition rates of the two cells were measured by CCK8 assay.(2)Divided the two kinds of cells into four groups: control(0 ?mol/L),MG132(5?mol/L),MPA(100 ?mol/L),MG132+MPA(5+100 ?mol/L),then all treated with the associated conditions for 24 h.a,Use CCK8 to detect the proliferation of the cells.b,To observe the morphological changes of the cells by inverted microscope.c,Use flow cytometry to detect the apoptosis of cells.d,Use immunohistochemistry to detect the expression of PR.e,Use westeron blot to detect the expression of PRA and PRB.Results:(1)The CCK8 results show: The viability(%)of HO-8910 cells treated with different concentration of MPA:(Role 24h)100.00�5.47,101.65�5.24,86.52�0.89,77.38�0.94,63.05�3.60;(Role 48h)100.00�0.94,104.45�0.37,71.06�3.22,55.70�0.74,48.08�0.08;(Role 72h)100.00�0.38,105.52�2.15,58.63�0.21,43.30�0.57,29.43�0.39;The viability(%)of SKOV3 cells treated with different concentration of MPA:(Role 24h)100.00�3.46,101.53�3.3,95.39�2.02,89.15�2.51,84.49�2.29;(Role 48h)100.00�2.70,99.58�0.29,89.22�1.67,86.37�0.22,78.44�5.08;(Role 72h)100.00�3.59,103.22�6.12,87.21�3.32,78.68�3.30,65.56�0.79.The viability of HO-8910 treated with MPA was higher than that of the SKOV3 cells,both of them were presenting with obviously dose-dependent and time-dependent cytotoxicity.The viability(%)of HO-8910 cells treated with different concentration of MG132:(Role 24h)100.00�1.30,98.73�1.09,83.29�0.43,71.36�3.10,66.54�8.33;(Role 48h)100�0.06,89.35�2.05,73.39�1.86,59.46�0.69,47.40�0.98;(Role 72h)100�2.70,79.67�2.80,54.72�3.56,48.42�0.20,34.90�0.25.The viability(%)of SKOV3 cells treated with different concentration of MG132:(Role 24h)100.00�4.58,94.39�4.33,86.12�3.18,75.73�2.57,61.41 �1.11;(Role 48h)100.00�3.40,80.18�2.73,62.11�3.89,51.76�3.99,46.97�1.75;(Role 72h)100.00�0.35,62.86�8.79,62.86�8.79,43.39�2.10,37.36�0.36,29.18�1.62.The viability of the two cells treated with MG132 were similarity,both of them were presenting with obviously dose-dependent and time-dependent cytotoxicity.The viability of SKOV3 cells treated with MG132 and MPA was obviously increased than that of MPA alone(p<0.05),while the cell viability of HO-8910 cells treated with MG132 and MPA has no significant difference with that of MPA alone(p>0.05).(2)The cell morphology observation results show: Cells in the control group were characterized by typical epithelioid cells,grow up like slabstone,were close together and were mainly roundness or long spindle in shape.In MG132 group,the number of cells decreased,the intercellular space increased,and there were more floating cells in the visual field,some of the cells became larger,and there were vacuoles in the cytoplasm.In MG132+MPA group,the number of cells decreased significantly,there were a large number of floating cells in the visual field and more vacuoles in the cytoplasm.There are significant differences in MPA group between the two cells,HO-8910 cells in MPA group were similar with the MG132 group,but the number of SKOV3 cells in MPA group were slightly decreased,arranged compactly,some of the cells stretch out spurius and were polygonal,no obvious floating cells were found in cytoplasm.(3)The flow cytometry results show: The cell apoptosis rate(%)of HO-8910 cells in each group: the control group was 0.46�0.04,the MG132 group was 7.43�0.83,the MPA group was 11.80�0.52,and the MG132 + MPA group was 12.36�0.42.The cell apptosis rate(%)of SKOV3 cells in each group: the ontrol group was 0.64�0.06,the MG132 group was 7.65�0.41,the MPA group was 6.86�0.54 and the MG132+MPA group was14.56�0.65.The apoptosis rate of SKOV3 cells in MG132+MPA group was significantly higher than that of MPA alone group(P<0.05),while in HO-8910 cells,there was no sinificant difference between the apoptosis rate of MG132+MPA group and MPA alone group(P>0.05).(4)The results of immunohistochemistry showed that the expression of PR was abundant in cell nucleus,and the cytoplasm was also expressed.The expression of PR in HO-8910 cells was higher than that of SKOV3 cells.The expression of PR in groups of MG132 and MG132+MPA were higher than that of group control(P<0.05),while there is no difference between the group MPA and group control(P>0.05).(5)The results of western blot showed that the expression of PRB in HO-8910 and SKOV3 cells was significantly higher than that of PRA.The expression of PRB and PRA in MG132 group and MG132+MPA group were higher than those in group control(P<0.05),while there is no difference between the group MPA and group control(P >0.05).Conclusion:(1)The proliferation inhibition and apoptosis indution effect of MPA on PR high expression cell line HO-8910 was stronger than that of PR low expression cell line SKOV3.(2)Proteasome inhibitor MG132 can inhibit the proliferation and induce apoptosis of HO-8910 and SKOV3,and also enhance the sensitivity of MPA to the proliferation inhibition and apoptosis induction of SKOV3 cells.(3)Proteasome inhibitor MG132 combined with MPA can promte the expression of PR in cells.This may be related to the phenomenon of MG132 increasing the sensitivity of SKOV3 cells to MPA.