Objective:

2

To investigate the anti-tumor effects of cinobufacini (CINO) on hepatocellular carcinoma (HCC) induced by des-gamma-carboxy-prothrombin (DCP) and to uncover the underlying mechanisms.

Methods:

2

The inhibitory effect of CINO on HCC cell proliferation was evaluated using the cell counting kit-8 method

and the apoptosis rate was quantified using flow cytometry. Immunofluorescence and Western blot analyses were used to investigate the differential expression of proteins associated with cell growth

apoptosis

migration

and invasion pathways after CINO treatment. The therapeutic potential of CINO for HCC was confirmed

and the possibility of combining cinobufacini with c-Met inhibitor for the treatment of primary HCC was further validated by

in vivo

experiments.

Results:

2

Under the induction of DCP

CINO inhibited the activity of HCC cells

induced apoptosis

and inhibited migration and invasion. Upon the induction of DCP

CINO regulated c-Met activation and the activation of the phosphatidylinositol-3 kinase/protein kinase B (PI3K/AKT) and mitogen-activated protein kinase kinase/extracellular signal-regulated kinase (MEK/ERK) pathways. In a mouse model of HCC

CINO exhibited significant antitumor effects by inhibiting the phosphorylation of c-Met and the downstream PI3K/AKT and MEK/ERK pathways in tumor tissues.

Conclusions:

2

CINO inhibited HCC cell growth

promoted apoptosis

and suppressed HCC cell invasion and migration by targeting c-Met and PI3K/AKT and MEK/ERK signaling pathways under DCP induction.