Research Of Mangiferin On Inhibiting The Proliferation And Invasion Of Human Ovarian Adenocarcinoma OVCAR8 Cells Via The Regulation Of YAP

Objective:Ovarian cancer is one of the most malignant gynecologic neoplasms in women and has the worst prognosis among female cancers based on their 5-year survival rates.Our previous study indicated that mangiferin possessed an anti-neoplastic effect on human ovarian cancer cells by targeting Notch3.Besides,it has been demonstrated that Notch signaling is functionally important downstream effector of YAP,therefore we hypothesized that YAP might be involved in the anti-tumor effect of mangiferin.Herein,the current study was aimed to investigate the effect of mangiferin on the human ovarian cancer OVCAR8 cell proliferation,apoptosis,migration and invasion by the regulation of YAP,further revealing the molecular anti-cancer mechanism of mangiferin in ovarian cancer treatment and providing experimental basis for the development of mangiferin as a novel and efficient antitumor drug in the treatment of ovarian cancer.Method:(1)The effect of mangiferin on YAP expression in OVCAR8 cells was detected by western blot.(2)YAP overexpression plasmids were constructed artificially.The experimental group was transfected with YAP overexpression plasmids and then cultured with mangiferin.The control group was transfected with empty vector and then cultured with mangiferin.(3)The cell proliferation of experimental group and control group were detected by MTT colorimetric assay,phase contrast microscope,Hoechst 33342 staining and cell colony formation assay,in order to investigate the effect of mangiferin on the cell proliferation through the regulation of YAP.(4)The expression of mitochondrial apoptosis-related proteins and the apoptosis in the experimental group and control group were respectively detected by western blot and Annexin V-FITC/PI flow cytometry,in order to explore the effect of mangiferin on the apoptosis through the regulation of YAP.(5)The cell migration and invasion ability of experimental group and control group were respectively detected by wound healing assay and Matrigel cell invasion assay,in order to investigate the effect of mangiferin on the cell migration and invasion through the regulation of YAP.Result:(1)YAP expression in OVCAR8 cells was decreased significantly after mangiferin treatment.(2)The inhibitory ratio of experimental group at 12 h,24h,36 h and 48 h after mangiferin treatment were significantly lower than that of control group,respectively(P<0.01).Control group cells died a lot and nuclear fragmentations were highly agglutinated in bright blue,whereas experimental group had significantly more viable cells and the nuclei were all pale blue and round.The number of cell colonies in the experimental group was significantly higher than that in the control group(P<0.01).(3)The expression of cleaved caspase-3 and cleaved PARP in the control group were significantly higher than that of experimental group.The number of early and late apoptotic cells in the control group were significantly higher than those in the experimental group,and the total apoptotic percentage of control group was significantly higher than that of experimental group(P<0.05).(4)Cell scratch fusion was observed in the experimental group 24 h after mangiferin treatment,while cell scratch in the control group was still clearly visible,with the significant difference in fold increase(P<0.01).Matrigel cell invasion assay results showed that the difference of fold increase between experimental group and control group was statistically significant(P<0.05).Conclusion:Mangiferin can effectively inhibit the expression of YAP in human ovarian cancer OVCAR8 cells.Besides,mangiferin can inhibit the OVCAR8 cell proliferiation,migration and invasion,and induce apoptosis through the downregulation of YAP.