Study on anti-hepatoma effect of ginsenoside Rh2 and mechanism of degradation of β-catenin through activating GSK-β / 中草药

ABSTRACT

Objective: To investigate the anticancer effect and the mechanism of ginsenoside Rh2 on animal model of HepG2 liver carcinoma. Methods: HE staining was used to observe cell morphology. Immunohistochemical staining was used to detect the expression of GSK-3β, β-catenin, and MMP-3 in isolated single cells. The activity of GSK-3β was checked by ELISA kit. The expression levels of GSK-3β, β-catenin, Bax, Bcl-2, CyclinD1, and MMP-3 genes were measured by qRT-PCR. The expression of β-catenin and GSK-3β proteins were determined by Western blotting. Results: HE staining showed that the nucleus was atypia and account for a large proportion of the whole cell in HepG2 group and HepG2-β-catenin group. But nucleus atypia in HepG2-β-catenin group was more obvious. Condensation nuclei and a lot of broken cells were observed in HepG2-β-catenin + ginsenoside Rh2 group and HepG2 + ginsenoside Rh2 group. However, condensation nuclei and broken cells in HepG2 + ginsenoside Rh2 group were more obvious. Immunohistochemical results indicated the expression of GSK-3β increased, while β-catenin and MMP-3 expression decreased in HepG2-β-catenin + ginsenoside Rh2 group, compared with HepG2-β-catenin group. The expression of β-catenin and MMP-3 in HepG2 + ginsenoside Rh2 group was lower than that in HepG2-β-catenin + ginsenoside Rh2 group, while GSK-3β was no significant difference. The ELISA results indicated that the activity of GSK-3β was increased in HepG2 + ginsenoside Rh2 group and HepG2-β-catenin + ginsenoside Rh2 group. Compared with HepG2-β-catenin + ginsenoside Rh2 group, the expression of Bax gene in HepG2 + ginsenoside Rh2 group increased significantly, and the expression levels of Bcl-2, CycliD1, and MMP-3 genes were also significantly lower, the difference was statistically significant (P <0.01). The Western blotting results showed that compared with HepG2-β-catenin + ginsenoside Rh2 group, the expression of β-catenin protein in HepG2 + ginsenoside Rh2 group was also significantly lower, the difference was statistically significant (P < 0.01). Conclusion: In vivo experiment shows that weight of tumor is decreased by ginsenoside Rh2 through activating GSK-3β to degrade β-catenin and could inhibit the ability of HepG2 cells metastasis.