The Effect Of Selenium Capparis Spinosa Polysaccharide On Cytoskeleton Protein RhoA And Cdc42 Of HepG2 Cells

The study of modern pharmacology shows that selenium Capparis spinosa polysaccharides(Se-CSPS)has the functions of anti-tumor,anti-oxidation,anti-virus,heavy metal resistance and immunoregulation.The pharmacological activity of Se-CSPS is more significant than polysaccharide and selenium.Se-CSPS has low side effect and is easily absorbed and utilized by the body,especially in anti-tumor,it can kill tumor cells directly and inhibit the proliferation of tumor cells.In recent years,there have many reports on Se-CSPS,but the mechanism of anti-tumor action of Se-CSPS are not reported.Therefore,in this study,we used vitro experiments to study the relationship between the cell cycle arrest and cytoskeleton protein RhoA and Cdc42 of HepG2 cells by Se-CSPS.It provides a theoretical support for Se-CSPS on HepG2 cells cycle arrest affecting.In this experiment,MTT method was used to detect the inhibitory effect of Se-CSPS on the proliferation of HepG2 cells.With the concentration of 0.064,0.128,0.256,0.512,1.024?mol·L-1 Se-CSPS administrated to the HepG2 cells for 72h.The growth inhibition rate was 17.15%,33.8%,53.47%,66.92%,79.64%.The inhibition rate increased gradually with the increase of the concentration by Se-CSPS,and it was dose dependent manner.The half inhibitory concentration(IC50)was 0.249?mol·L-1 through calculation after Se-CSPS administrated to the HepG2 cells for 72h.The results showed that Se-CSPS could inhibit the proliferation of HepG2 cells.By using flow cytometry and propidium iodide dye staining,the blank control group,the positive drug hydroxycamptothecin group 2.65?mol·L-1 and three different doses(0.125?0.25?0.5 ?mol·L-1)of Se-CSPS were administered to the HepG2 cell for 48h.The inhibition rate of G2/M period was 13.536%,65.533%,17.783%,22.036%and 39.236%,The inhibition rate increased gradually with the increase of the concentration by Se-CSPS,and it was dose dependent manner.The results showed that Se-CSPS in diffeerent doses could block HepG2 cells in G2/M period effectively after 48h.the blank control group,the positive drug hydroxycamptothecin group 2.65 ?mol·L-1 and different doses(0.125?0.25?0.5 ?mol·L-1)of Se-CSPS were used in HepG2 cells for 48h by using Western Blot staining.The expression of CDK1,CyclinB1,P21 and cytoskeletal protein RhoA and Cdc42 were analyzed.The result is:CDK1/?-actin:0.91,0.29,0.78,0.49,0.32.CyclinB1/?-actin:0.92,0.34,0.76,0.48,0.31.P21/?-actin:0.40,0.73,0.47,0.62,0.78.RhoA/?-actin:0.92,0.31,0.78,0.50,0.32.Cdc42/?-actin:0.90,0.36,0.76,0.54,0.32.The protein expression of CDK1,CyclinB1,RhoA and Cdc42 was significantly lower than the blank control group,with the increase of the concentration of Se-CSPS.The protein expression of P21 was significantly higher than that in the control group.The experimental results were statistically significant.The results showed that after Se-CSPS were used in HepG2 cells,the expression of CDK1,CyclinB1,RhoA and Cdc42 protein was significantly down,and the expression of P21 was significantly up.The results showed that Se-CSPS had proliferation inhibitory effect on HepG2 cells.Se-CSPS blocked cells in the G2/M period after 48h.After testing the expression of cell cycle related protein and cytoskeleton protein RhoA and Cdc42.The mechanism of HepG2 cells cycle arrest by Se-CSPS was as follows:(1)The expression of cyclin protein CDK1 and CyclinB1 was down regulated,the expression of P21 was up regulated and the activity of CDK1/CyclinB1 was reduced.(2)The expression of cytoskeleton protein RhoA and Cdc42 was downregulated.It affects the morphology of cytoskeleton,the polymerization state of actin cytoskeleton,the migration of cells,and the operation of cell cycle,so the HepG2 cells were blocked in the G2/M phase.