Ginsenoside inhibits aerobic glycolysis in hepatocellular carcinoma through the hsa-miR-139-5p/AURKA axis

Purpose: Ginsenoside (Rg3) is an important small-molecule with anti-tumor and immune-enhancing properties, mainly used in clinical Adjuvant chemotherapy for hepatocellular carcinoma (HCC). In this study, we explored the active targets and mechanisms of action of Rg3 in HCC to provide new basic data for clinical pharmaceutical research.

Methods: HepG-2 cells were treated with a concentration gradient of Rg3, cell activity was detected using the CCK-8 assay, and the IC50 was calculated. Flow cytometry was used to detect the apoptosis-inducing effect and the cyclic distribution of the drug in the cells. An enzyme-linked immunosorbent assay (ELISA) kit was used to detect lactic acid (LC) and glucose (GLU) contents after cell treatment. High-throughput data analysis was performed to explore HCC-associated glycolytic genes, analyze the correlation between biomarkers and clinical data, and confirm the gene-binding targets using a luciferase assay. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blot (WB) were utilized to assess gene and protein expression in clinical samples and cell subgroups. The reliability of the cellular experiments was verified in vivo in transplanted mouse tumors.

Results: Rg3 at a concentration gradient inhibited LC and GLU secretion and induced Apoptosis to inhibit cell proliferation. High-throughput data analysis revealed six biomarkers, among which miR-139-5p was highly expressed in tumor tissues and cells. Immunofluorescence experiments revealed targeted binding to Aurora Kinase A (AURKA). By targeting miR-139-5p, Rg3 inhibited tumor proliferation to reverse AURKA expression. Rg3 induced Apoptosis, reduced the expression levels of AURKA, GLUT1, and GLUT4 and increased the expression of P53. miRNA-inhibit and Overexpression of Aurora Kinase A (OE-AURKA) were combined to stimulate the expression of P53 proteins, which significantly reversed the glycolytic activation of miRNA-inhibit and OE-AURKA and induced Apoptosis. In vivo, Rg3 inhibited Apoptosis and the expression of glycolytic proteins by targeting miR-139-5p, consistent with the in vitro results.

Conclusion: Rg3 inhibits the proliferation of HepG-2 cells and explores a novel mechanism by which the miR-139-5p/AURKA axis suppresses aerobic glycolysis in HCC.

Aerobic glycolysis; Drug activity; Hepatocellular carcinoma; Rg3; miR-139-5p.