Abstract

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality worldwide, which is difficult to control with conventional surgical and chemotherapeutic approaches owing to distant metastasis. Therefore, alternative medicines with high efficacy but minimal side effects are urgently needed. Recently, increasing evidence has demonstrated that ononin, an isoflavone glycoside widely distributed in herbs and food plants, exhibits potent anticancer effects across various cancers. Our research aimed to determine the influence of ononin on HCC growth and metastasis and identify the potential mechanism of action. The anti-proliferative and anti-metastatic properties of ononin in human HCC cells were determined using MTT, colony formation, flow cytometry, wound-healing, and Transwell assays. The expression of cell cycle-related proteins, matrix metalloproteinases (MMP-2 and MMP-9), and PI3K/AKT/mTOR pathway-related molecules in HCC cells was examined by western blotting. TGF-β1 or the PI3K activator 740Y-P was used for combined cell treatment with ononin to validate the effects of ononin on TGF-β1-triggered HCC cell metastasis and the role of the PI3K/AKT/mTOR pathway in ononin's anticancer effects. The in vivo therapeutic effects were finally validated in xenograft mouse models. Ononin suppressed HCC cell growth, induced G0/G1 cell cycle arrest, and reduced CDK6, cyclin D1, and CDK4 protein levels in HCC cells. Ononin inhibited HCC cell migration and invasion and downregulated MMP-2/9 expression with or without TGF-β1 stimulation. Ononin attenuated p-PI3K, p-AKT, and p-mTOR protein levels in HCC cells, and 740Y-P abolished the repression of ononin on HCC cell growth and metastasis. Additionally, ononin restricted tumor growth, decreased Ki67 and p-AKT expression, and curbed lung metastasis in xenograft mouse models. Ononin inhibits HCC growth and metastasis by downregulating MMP-2 and MMP-9 through inactivating the PI3K/AKT/mTOR signaling.