Study On The Mechanism Of Apoptosis Of Human Ovarian Cancer HO-8910 Cells By Fuzheng Kangai Tang-containing Serum Combined With Cisplati

Objective: To explore the effect and mechanism of Fuzheng Kangai Decoction combined with cisplatin on apoptosis of ovarian cancer HO-8910 cells at cellular level.To provide experimental basis for Fuzheng Kangai Decoction combined with cisplatin in the treatment of ovarian cancer.Materials and methods: 1.To observe the effects of serum containing Fuzheng Kangai Decoction with different concentrations on the proliferation of human ovarian cancer HO-8910 cells.Preparation of Fuzheng Kangai Decoction medicated serum: 20 SPF SD rats(males)were randomly divided into two groups,10 rats in each group.After a week of adaptive feeding,they began to be given gastric lavage.After concentrating the purchased Fuzheng Kangai Decoction by 6 times,they were given gastric lavage by 9 times of the equivalent dose of human beings.Each rat in a group was given 4.05 ml of concentrated Fuzheng Kangai Decoction for 10 days,and marked as medicated serum group.The remaining group of rats were fed with 4.05 ml of normal saline for 10 days and marked as blank serum group.On the 10 th day of gastric administration,one hour after administration,chloral hydrate was injected intraperitoneally,blood was taken after anesthesia,and the medicated serum and blank serum were prepared and refrigerated for later use.Inoculate human ovarian cancer HO-8910 cells in a 96-well plate,then mix the serum containing Fuzheng Kangai Decoction with blank serum according to the ratio of 1: 0,1: 1 and 1: 3,and mark it as high concentration group,medium concentration group and low concentration group in turn,and then intervene in human ovarian cancer HO-8910 cells for 24 h,and detect by MTT(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide)method to observe the proliferation of human ovarian cancer HO-8910 cells with different concentrations of serum containing Fuzheng Kangai Decoction.2.To observe the effect of Fuzheng Kangai Decoction serum combined with cisplatin on human ovarian cancer HO-8910 cells.Human ovarian cancer HO-8910 cells were randomly divided into blank control group,Fuzheng Kangai Decoction containing serum combined with cisplatin group and cisplatingroup.According to MTT results,the optimal concentration of Fuzheng Kangai Decoction containing serum combined with cisplatin and cisplatin was selected to intervene human ovarian cancer HO-8910 cells respectively.The cell proliferation of each group was detected by CCK8 method.Cell apoptosis in each group was detected by flow cytometry.The expression of FAS(Apopotisis-1)and FAS-L(Fas ligand)in cells of each group was detected by immunochemistry.The expression of Caspase-3(Cysteinyl Aspartate Apecific Proteinase 3)and Caspase-8(Cysteinyl Aspartate Apecific Proteinase 8)protein in each group was detected by Western-blot.Results: 1.The effects of serum containing Fuzheng Kangai Decoction with different concentrations on the proliferation of human ovarian cancer HO-8910 cells.From the MTT results,it can be seen that the serum containing Fuzheng Kangai Decoction can inhibit the proliferation of human ovarian cancer HO-8910 cells,and the high concentration serum containing Fuzheng Kangai Decoction has the best inhibitory effect on human ovarian cancer HO-8910 cells(p<0.01).2.The effect of Fuzheng Kangai Decoction serum combined with cisplatin on human ovarian cancer HO-8910 cells.The results of CCK8 detection showed that compared with the blank control group,the OD value of cells in each group was significantly lower,with statistical significance(p<0.01),and the OD value of cells in Fuzheng Kangai Decoction combined with cisplatin group was also statistically significant(P < 0.01).The results of flow cytometry showed that compared with the blank control group,the apoptosis rate of all groups increased with statistical significance(p<0.01),and the apoptosis rate of Fuzheng Kangai Decoction combined with cisplatin group also had statistical significance(P < 0.01).The results of immunochemistry showed that,compared with the blank control group,the expression of FAS and FAS-L in cells of each group increased with statistical significance(p<0.01),and the expression of FAS and FAS-L in cells of Fuzheng Kangai Decoction combined with cisplatin group also showed statistical difference(P < 0.01).The results of Western-blot showed that compared with the blank control group,the protein expression of Caspase-3 and Caspase-8 in each groupincreased with statistical significance(p<0.01).The protein expression of Caspase-3 and Caspase-8 in Fuzheng Kangai Decoction combined with cisplatin group had no statistical difference(p>0.05),but the Caspase-8 in Fuzheng Kangai Decoction combined with cisplatin group had no statistical difference.Conclusion: 1.High concentration serum containing Fuzheng Kangai Decoction has the best inhibitory effect on the proliferation of human ovarian cancer HO-8910 cells.2.Fuzheng Kangai Decoction serum combined with cisplatin can inhibit the proliferation of human ovarian cancer HO-8910 cells and increase the apoptosis of HO-8910 cells.3.Fuzheng Kangai Decoction serum combined with cisplatin can activate FAS/FAS-L pathway by up-regulating the protein expression of Fas and Fas-L in HO-8910 cells,and further activate Caspase-3 and Caspase-8,thus promoting the apoptosis of HO-8910 cells.