The Effect Of Genistein To RWPE-1 Cells Proliferation Induced By Testosterone Under The Control Of IGF-1

Object:To study the independent effect and combined effect of genistein and IGF-1 to RWPE-1 cells proliferation induced by testosterone,and to observe the inhibition and the influence of cell cycles of genistein to cell proliferation under the suitable concentration of IGF-1.In addition,by using detection of cyclin B₁,cyclinD₁ and cyclin-dependent kinase 4 so as to provide a new idea for the study of molecular mechanism of genistein on cell proliferation.Methods:This experiment used the RWPE-1 cells which was promoted cell proliferation by testosterone propionate.MTT method was applied to detect the effect of IGF-1 and genistein on the proliferation of RWPE-1 cells,and to observe the inhibition of genistein to cell proliferation under the suitable concentration of IGF-1.The flow cytometry analysis was used to observe the effect of IGF-1 and genistein on the cycle of RWPE-1 cells.Western blot was used to study the protein expression levels of cyclin B₁,cyclinD₁ and cyclin-dependent kinase 4 was treated with IGF-1 and genistein.Results:(1) The experiment of MTT demonstrate that more and more obvious inhibition of cell proliferation occurred as the concentration of genistein increased,and 40.00μmol/L of genistein,3 days was the suitable concentration and observation time respectively.It also demonstrate that more and more obvious promotion of cell proliferation appeared as the concentration of IGF-1 increased,and 6.4nmol/L of IGF-1,3 days was the suitable concentration and observation time respectively.The cell proliferation which was treated with genistein under the suitable concentration of IGF-1 was obviously inhibited and the degree was significantly higher than that in cell proliferation treated only with genistein.(2) The experiment of flow cytometry analysis demonstrate that compared with the control group,the cell percentage which was treated with genistein under the suitable concentration of IGF-1 in G₁ phase decreased and in both S phase and G₂/M phase increased.(3) The semi-quantitative analysis which was used in Western-Blot showed that the expression of the cyclinD₁,CDK4 decreased and the expression of cyclinB₁ increased when the cells were treated with genistein alone,the expression of the cyclinB₁,cyclinD₁,CDK4 all increased with IGF-1,the expression of the cyclinD₁,CDK4 decreased and the expression of cyclinB₁ increased when the cells were treated with genistein under the suitable concentration of IGF-1.Conclusion:(1) The genistein can inhibit the proliferation of RWPE-1 cells and the more and more obvious inhibition occurred as the concentration of genistein increased and the time prolonged.The inhibition of cell proliferation which was treated with genistein under the suitable concentration of IGF-1 was more obvious than that in cell proliferation treated only with genistein.(2) The inhibition of cell proliferation which was treated with genistein is probably related to the blocking of G₂/M phase and the IGF-1 can promote the G₂/M phase block effect of genistein on RWPE-1 cells proliferation.(3) The genistein can inhibition the proliferation of RWPE-1 cells under the control of IGF-1 and it showed that the influence was related to the expression of the cyclinB₁,cyclinD₁,CDK4.