The Effects Of 6-shogaol On Cisplatin Triggering Apoptosis In SKOV-3 And A2780 Cells

Background and objectives:Microtubules are important components of the cytoskeleton.Microtubules play a crucial role in cell mitosis.Therefore,the study of tubulin has become an important target for the development of anticancer drugs.Based on platinum combined with Paclitaxel chemotherapy is the first-line chemotherapy for ovarian cancer.However,the resistance of platinum and paclitaxel is a serious constraint on the prognosis of ovarian cancer patients.Recent research reports,6-shogaol induced microtubule damage and inhibited microtubule polymerization.The objective of this study is to investigate the effects of 6-shogaol on cisplatin triggering apoptosis in SKOV-3 and A2780 cells and 6-shogaol leading to microtubule damage in order to provide a basis for the treatment of ovarian cancer with platinum combined with 6-shogaol.Methods:1?The inhibitory effect of 6-shogaol,paclitaxel,cisplatin and 6-shogaol combined with platinum,paclitaxel combined with platinum on the growth of human ovarian cancer A2780,SKOV-3 cells were investigated by using crystal violet assay.2?The effects of 6-shogaol,Paclitaxel,cisplatin and 6-shogaol combined with platinum,paclitaxel combined with platinum on cell cycle distribution and cells apop-tosis rate of human ovarian cancer A2780 and SKOV-3 cells were detected by using flowcy-tometry.3?Immunofluorescence staining was used to observe mitosis of A2780 and SKOV-3 cells treated with 6-shogaol and Paclitaxel and to calculate the percentage of M phase.Results:1?6-shogaol,Paclitaxel and cisplatin inhibited the growth of A2780 and SKOV-3cells in a concentration dependent manner.The half inhibitory concentrations of6-shogaol,Paclitaxel and cisplatin induced A2780 and SKOV-3 cells after 72 hours were15.277±0.380?mol/L and15.540±0.230?mol/L,0.018±0.001?mol/L and0.023±0.002?mol/L,6.983±0.538?mol/L and 6.177±0.440?mol/L.The half inhibitory concentrations of 6-shogaol(IC₁₀)combined with different concentration of cisplatin,cisplatin(IC₁₀)combined with different concentration of 6-shogaol,paclitaxel(IC₁₀)combined with different concentration of cisplatin and cisplatin(IC₁₀)combined with different con-centrations of paclitaxel induced A2780 and SKOV-3cells after 72 hours were 2.913±0.187?mol/L and 1.954±0.136?mol/L,3.212±0.225?mol/L and 3.352±0.258?mol/L,2.768±0.173?mol/L and 2.388±0.203?mol/L,0.003±0.0001?mol/L and 0.004±0.0001?mol/L.Results:compared with 6-shogaol combined with cisplatin and cisplatin,cell growth inhibition was significantly increased?P<0.05?and 6-shogaol significantly improved the growth inhibitory effect of cisplatin on ovarian cancer cells.compared with cisplatin combined with 6-shogaol and 6-shogaol,cell growth inhibition was significantly increased?P<0.05?and cisplatin significantly increased the growth inhibitory effect of 6-shogaol on ovarian cancer cell.There was no significant difference in cell growth inhibition incubated by6-shogaol combined with cisplatin and paclitaxel combined with cisplatin?P>0.05?.There was no significant difference in growth inhibition of A2780 and SKOV-3 cells incubated by cisplatin(IC₁₀)combined with different concentrations of 6-shogaol and6-shogaol?P>0.05?.The inhibitory effect of 6-shogaol(IC₁₀)combined with different concentrations of cisplatin and cisplatin on the growth inhibition of SKOV-3cells was significantly better than that of A2780 cells?P<0.05?.2?The A2780 and SKOV-3 cells incubated with 6-shogaol and paclitaxel were arrested in G2/M phase in concentration-dependence manner.A2780 and SKOV-3cellsincubatedwithCisplatinwereblockedinG1/Sphasein concentration-dependence manner.6-shogaol(IC₁₀)combined with cisplatin(IC₅₀)and paclitaxel(IC₁₀)combined with cisplatin(IC₅₀)were able to block the cell cycle in the G1/S phase.Cisplatin(IC₁₀)combined with 6-shogaol(IC₅₀)and cisplatin(IC₁₀)combined with paclitaxel(IC₅₀)can block the cell cycle in the G2/M phase.6-shogaol,paclitaxel,cisplatin and combined groups respectively increased the apoptosis of A2780 and SKOV-3 cells in a time-dependent manner.Results:There was no significant difference between 6-shogaol and paclitaxel in promoting apoptosis of A2780 cells?P>0.05?.In the promotion of apoptosis,the effect of 6-shogaol was higher than paclitaxel in SKOV-3 cells.6-shogaol(IC₁₀)combined with cisplatin induced apoptosis was significantly better than that of cisplatin group?P<0.05?.3?The percentage of M phase of ovarian cancer cells incubated by 6-shogaol and paclitaxel were respectively in A2780?32.79%and 33.16%?and SKOV-3?32.65%and 32.77%?.The percentage of M phase of ovarian cancer cells in the control group was in A2780?5.27%?and SKOV-3?4.71%?.Results:Compared with the control group,the percentage of mitotic cells of 6-shogaol and paclitaxel were significantly different?P<0.05?.There was no significant difference in the percentage of mitotic cells incubated by 6-shogaol and paclitaxel in ovarian cancer cells?P>0.05?.Compa-red with the control group,the green fluorescence brightness of microtubules of hum-an ovarian cancer A2780?SKOV-3 cells is not obvious by incubated with 6-shogaol for 24 hours.The microtubules were broken and had cavities in cytoplasm.Microtubu-les are no longer characterized by typical characteristics of radial distribution,and there are multinucleated cells in A2780 and SKOV-3 cells.Conclusions:1?The growth of human ovarian cancer A2780?SKOV-3 cells was inhibited by6-shogaol in vitro.2?6-shogaol inhibited the polymerization of microtubules and increased the inhibitory effect of cisplatin on human ovarian cancer A2780 and SKOV-3 cells,which blocked the cell cycle in G2/M phase and promoted the apoptosis of ovarian cancer cells.3?For MyD88 positive SKOV-3 cells induced taxol resistance and tubulin dependent paclitaxel resistance,6-shogaol can be used as an alternative to paclitaxel in the treatment of ovarian cancer,which was of prospective significance.