Effect and mechanism of gallic acid on the proliferation, migration, invasion, and apoptosis of human hepatocellular carcinoma HepG2 cells
Journal Title: Journal of Clinical Hepatology - Year 2025, Vol 41, Issue 3
Abstract
[Objective] To investigate the effect of gallic acid (GA) on the proliferation, migration, invasion, and apoptosis of human hepatocellular carcinoma HepG2 cells and its mechanism. [Methods] HepG2 cells were treated with different concentrations of GA (0, 5, 10, 20, 30, 40, and 50 μg/mL) for 24 and 48 hours, and CCK8 assay was used to measure cell viability and calculate IC50. The experiment was divided into control group (HepG2 cells), 5 μg/mL GA group, 10 μg/mL GA group, and 20 μg/mL GA group. Plate colony formation assay was used to evaluate the effect of GA on cell proliferation; wound healing assay and Transwell chamber assay were used to observe the effect of GA on cell migration and invasion; flow cytometry was used to observe the effect of GA on cell apoptosis; Western blot was used to measure the expression of matrix metallopeptidase-2 (MMP-2), matrix metallopeptidase-9 (MMP-9), and apoptosis-related proteins. A one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. [Results] The mean IC50 value of GA on HepG2 cells was 38.02±2.58 μg/mL at 24 hours and 18.36±1.54 μg/mL at 48 hours. The number of cell colonies was 239.00±29.45 in the control group, 210.00±19.00 in the 5 μg/mL GA group, 144.33±16.03 in the 10 μg/mL GA group, and 57.00±9.55 in the 20 μg/mL GA group, suggesting that compared with the control group, each GA group had a significant reduction in cell colony formation ability (all P<0.05). After 24 hours of treatment, the cell migration rate was 42.62%± 7.82% in the control group, 35.34%±6.42% in the 5 μg/mL GA group, 21.85%±4.42% in the 10 μg/mL GA group, and 12.57%± 3.54% in the 20 μg/mL GA group, respectively, in these four groups, and the number of transmembrane cells in these four groups was 230.30±15.30, 182.12±12.60, 137.20±7.50, and 124.40±6.80, respectively, suggesting that compared with the control group, each GA group had significant reductions in migration rate and the number of transmembrane cells (all P<0.05). After 48 hours of treatment, the cell apoptotic rate was 0.67%±0.08% in the control group, 13.27%±1.07% in the 5 μg/mL GA group, 20.94%± 2.45% in the 10 μg/mL GA group, and 40.74%±2.63% in the 20 μg/mL GA group, and compared with the control group, each GA group had a significant increase in cell apoptosis rate (all P<0.05). Compared with the control group, each GA group had significant reductions in the protein expression levels of MMP-2 and MMP-9 (all P<0.05) and significant increases in the protein expression levels of Bax and cleaved caspase-3 (all P<0.05). [Conclusion] GA can inhibit the proliferation, migration, and invasion of HepG2 cells and promote the apoptosis of HepG2 cells, possibly by regulating MMP-2, MMP-9, and the apoptosis-related proteins Bax/Bcl-2.
Authors and Affiliations
Zhiru WANG, Wenjing ZHAO, Xi CHEN
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