| Ovarian cancer is a very deadly gynecological malignancy.Most patients are diagnosed at an advanced stage due to the lack of specific symptoms in the early stage and screening methods,which poses a huge threat to women’s life and health.Currently carboplatin(CBP)and paclitaxel(PTX)are used for first-line chemotherapy in ovarian cancer.In view of the toxic and side effects of CBP/PTX therapy and the problem of CBP-resistance,this paper focuses on the synergistic antitumor effect of the combination of apigenin(API)and CBP/PTX on Caov-3 cells,and the reversal of resistance of ovarian cancer cells Hey A8 to CBP.The research results and conclusions are as follows:(1)The MTT method was used to detect the effect of the combination of API and CBP/PTXon the proliferation of Caov-3 cells.It was found that API significantly enhanced the inhibitory effect of CBP/PTX on the proliferation of Caov-3 when the combined effect was 48 h,and the combination index was 0.54-0.98.The results of PI single staining,AV/PI double staining,and real-time quantitative PCR analysis showed that the combination of API and CBP/PTX blocked more cells in S phase and G2 phase,and the expression levels of Bcl2/Bax ratio was significantly decreased.The combination of API and CBP/PTX induced an increase in scratch width and a decrease in the m RNA expression of MMP1,3,and 9.In the Caov-3nude mouse xenograft model,the tumor inhibition rateof 5 mg/kg API and 5 mg/kg CBP / 3mg/kg PTX groups(77.80%)was higher than that of 15 mg/kg CBP / 6 mg/kg PTX group(74.26%),and the doses of CBP and PTX decreased by 3 and 2 folds,respectively.The results of HE staining showed that the combination therapy did not cause damage to the main internal organs of nude mice.Immunohistochemical analysis further demonstrated that the combination of API and CBP/PTX down-regulated the expression of Bax/Bcl2.(2)Compared with the Caov-3 cell line,Hey A8 cells were CBP-resistant cell line with a drug resistance index of 13.35.The inhibitory effect of combination of API and CBP/PTX on the proliferation of Hey A8 cells was analyzed by MTT method,it was found that API could reverse the drug resistance of Hey A8 cells to CBP,and the resistance index of Hey A8 cells to CBP decreased to 4.07.The differentially expressed genes caused by the combination therapy were analyzed by transcriptomics and verified by the fluorescence real-time quantitative PCR,indicated that the combination of API and CBP/PTX reversed the drug resistance to CBP by reducing the expression of EGF to decrease the accumulation of STAT5 A in the nucleus.Thus,the cell cycle was arrested in S phase,and more cells were undergone apoptosis by CBP,which was the same as the results of PI single-staining and AV/PI double-staining.In the Hey A8 nude mouse xenograft model,the tumor inhibition rate of 5 mg/kg API and 10 mg/kg CBP / 3 mg/kg PTX group(83.22%)was higher than that of 50 mg/kg CBP / 6 mg/kg PTX group(82.75%),and the dose of CBP decreased 5 folds.The results of HE staining showed that the combination therapy did not cause damage to the main internal organs of nude mice.In summary,it was found that there was a synergistic effect between API and CBP/PTX,and the mechanism of anti-tumor effect of their combination has been revealed.Besides,the addition of API increased the sensitivity of Hey A8 cells to CBP,revealing the mechanism of drug resistance to CBP of Hey A8 cells.Moreover,the combination of API and CBP/PTX reduced the migration ability of tumor cells,limiting the malignant development of tumors,which has a great significance to the prognosis of patients.The pharmacodynamic evaluation in vivo proved that the combination of API and CBP/PTX has a significantly synergistic antitumor effect and reversal of CBP-resistance. |
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