Ginsenoside Rh2 and luteolin synergistically induce cellular senescence to suppress hepatocellular carcinoma progression through oxidative stress-mediated mechanisms.

BACKGROUND

Hepatocellular carcinoma (HCC) remains one of the leading causes of cancer-related mortality worldwide, and effective therapeutic options are still limited. Inducing cellular senescence has emerged as a promising strategy to restrain tumor progression. Ginsenoside Rh2 (Rh2) and luteolin (Lut) each exhibits anticancer activity; however, their behavior in combination and underlying mechanisms in HCC remain largely unexplored.

OBJECTIVE

To investigate the effects and molecular mechanisms of the Rh2/Lut combination in inducing cellular senescence in HCC.

METHODS

Cell proliferation, migration, and invasion were assessed using cell counting kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), colony formation, wound-healing, and Transwell assays. Synergy was evaluated using SynergyFinder. Cellular senescence and mitochondrial dysfunction were examined through SA-β-Gal staining, senescence marker detection, ROS measurement, flow cytometry, and mitochondrial membrane potential (MMP) assays. Mechanistic studies were conducted via western blotting (WB), immunofluorescence (IF), and rescue experiments with the Nrf2 activator sulforaphane (SFN). γH2AX staining was performed to assess DNA damage. In vivo antitumor efficacy and biosafety were evaluated in a nude-mouse xenograft model, including histopathological examination of major organs (H&E staining).

RESULTS

The Rh2/Lut (1:2) combination displayed strong inhibition of HCC cell proliferation, migration, and invasion. Mechanistically, the combination suppressed the Nrf2/HO-1 pathway while activating the p53/p21 signaling axis. Increased γH2AX expression indicated DNA damage-associated senescence. These effects were reversed by SFN treatment. In vivo, Rh2/Lut significantly inhibited tumor growth without inducing pathological abnormalities in major organs.

CONCLUSION

The Rh2/Lut combination triggers mitochondrial dysfunction-associated DNA damage and senescence through modulation of the Nrf2/HO-1-p53/p21 axis, exerting potent antitumor activity with favorable biosafety. These findings support Rh2/Lut as a promising therapeutic strategy for HCC.