Study Of The Effect Of Apatinib On Cisplatin-resistant Cells In Ovarian Cancer And Its Mechanism

Objective:To investigate whether apatinib affects the growth of human ovarian cancer cisplatinresistant cell line SKOV3/DDP and its responsiveness to cisplatin,and to explore its possible mechanisms,so as to provide new directions for the treatment of cisplatin-resistant patients with ovarian cancer.Methods:1.Human ovarian cancer cell line SKOV3 and human ovarian cancer cisplatin-resistant cell line SKOV3/DDP were selected as subjects,and the two cells were simultaneously treated with different concentrations of cisplatin for 24 h,48h and 72 h.The effect of cisplatin on the proliferation rate of these two kinds of cells was detected by CCK8 method,and IC50 and drug resistance multiple were calculated respectively.2.SKOV3/DDP was used as the main body of the study,and the cell proliferation rate was detected by CCK8 method after 24 h,48h and 72 h treatment with different concentrations of Apatinib,and IC50 was calculated.3.SKOV3/DDP was taken as the main body of the study and divided into four groups:blank control group,cisplatin group,Apatinib group and Apatinib + cisplatin group.Different drugs were given to each group for treatment,and the cell proliferation rate was detected and compared by CCK8 method.4.Western blot was used to detect and compare the expression differences of γH2AX,BRCA1 and Rad51 proteins in SKOV3/DDP cells divided into blank control group,cisplatin group and Apatinib + cisplatin group.5.Statistical software SPSS 26.0 and Graph Pad Prism 8.0 were used for data analysis and plotting.Quantitative data through the mean ± standard deviation((?)±s),comparison between the two groups using t test,using one-way ANOVA is compared between multiple sets of single factor analysis of variance,and LSD-t method was used for pairwise comparison,set P<0.05 was the test standard.Results:1.CCK-8 results showed that the cell proliferation rate of SKOV3/DDP was greater than that of SKOV3 at different concentrations of cisplatin at 24 h,48h and 72 h on SKOV3 and SKOV3/DDP cells(P < 0.05).2.The CCK-8 results showed that the cell proliferation rate was statistically significantly lower than that of the control group after the effective drug concentration of apatinib exceeded 32μmol/L at drug treatment times of 24 h,48 h,and 72 h(P < 0.05).Under the conditions of the same dosing time of apatinib,the proliferation rate of cells after the effective concentration of the drug exceeded 32μmol/L gradually decreased with the increase of the effective concentration of the drug(P < 0.05);under the conditions of drug intervention with the effective concentration of apatinib of 32μmol/L and 64μmol/L,the longer the drug treatment time,the lower the proliferation rate of cells(P < 0.05)3.The results of CCK-8 showed that the cell proliferation rate of Apatinib alone group was significantly decreased compared with control group(P < 0.05).The proliferation rate of cisplatin combined with Apatinib group was significantly lower than that of cisplatin monotherapy group(P < 0.05).The cell proliferation rate of cisplatin combined with Apatinib group was also significantly decreased compared with Apatinib monotherapy group(P < 0.05).4.Western blot results showed that compared with cisplatin monotherapy group,γH2AX protein levels in cisplatin combined Apatinib group were increased(P < 0.05),while BRCA1 and Rad51 protein levels were decreased(P < 0.05).Conclusion:In this study,we confirmed that Apatinib can inhibit the proliferation of human ovarian cancer cisplatin-resistant cell line SKOV3/DDP by CCK8 method,and has dose-effect dependence when the concentration is greater than 32μmol/L.At the same time,Apatinib can enhance SKOV3/DDP sensitivity to cisplatin,reverse cisplatin resistance,and has synergistic enhancement effect with cisplatin.The mechanism may be that Apatinib upregulates the expression of DNA damage-related protein γH2AX and down-regulates the expression of homologous recombination repair(HR)related proteins BRCA1 and Rad51,thus inhibiting the proliferation of cisplatin resistant cells,improving the sensitivity of cells to cisplatin,and reversing the resistance of ovarian cancer cells to cisplatin to a certain extent.It provides a new idea for the treatment of platinum-resistant ovarian cancer patients.