| Objective:Effect of betulinic acid on YAP protein in Hippo pathway and biological behavior of human hepatoma HepG2 cells.Methods:After betulinic acid was applied to human hepatoma HepG2 cells,the expression of YAP protein and YAP mRNA level in HepG2 cells were detected by western blotting and RT-PCR respectively.The accumulation of YAP protein in nucleus and cytoplasm was detected by immunofluorescence confocal.After YAP plasmid was transfected into HepG2 cells,the effect of betulinic acid on the biological behavior of HepG2 cells was detected by MTT assay,cell scratch assay,and cell colony formation assay,respectively.The effect of betulinic acid on human hepatocellular carcinoma HepG2 cells in vivo was detected by xenograft in nude mice.The expression of YAP protein in tumor tissues was detected by frozen section and immunofluorescence technique.Results:After treatment with betulinic acid,the level of YAP protein in HepG2 cells decreased in a concentration-dependent manner and YAP mRNA levels unchanged.Western blotting and immunofluorescence confocal results confirmed that betulinic acid could inhibit the nuclear accumulation of YAP protein.Moreover,betulinic acid inhibited cell proliferation,migration in a concentration-and time-dependent manner,and also inhibited cell colony formation in a concentration-dependent manner.The high expression of YAP reversed the inhibitory effect of betulinic acid on YAP protein and cell biological behavior including cell proliferation,migration,and colony formation capacity.The experiments in vivo also showed that compared with the control group,the tumor volume and weight decreased after betulinic acid treatment,and the expression of YAP protein in tumor tissues decreased.Conclusion:These findings revealed that betulinic acid inhibited cell proliferation,migration and colony formation by inhibiting YAP expression and nuclear localization.And,betulinic acid also inhibited the growth of xenograft in nude mice. |
PDF Full Text Request