[Study on Autophagy and Apoptosis Induced by Amiodarone Combined with Glycyrrhetinic Acid in HepG2 Cells]

Abstract

Objective: To investigate the effects of amiodarone combined with glycyrrhetinic acid on the activity, apoptosis and autophagy in human hepatoma HepG2 cells.

Methods: After using amiodarone and glycyrrhetinic acid alone or in combination treatment for HepG2 cells, MTT assay was used to detect cell proliferation, Annexin Ⅴ/PI flow cytometry was used to detect apoptosis; Western blot was used to detect the expression of autophagy-related proteins Beclin-1, LC3 and p62. The formation of EGFP-LC3 green fluorescent aggregates was observed under a fluorescence microscope. The effects of autophagy on cell proliferation and apoptosis were studied by autophagy inhibitor hydroxychloroquine (HCQ) and autophagy promoter Rapamycin.

Results: The cell viability in combination group was lower than that in single drug group, and the apoptosis rate was higher than that in single drug group. Compared with single-drug group, the expressions of Beclin-1 and LC3Ⅱ protein in the combination group were higher than that in the single-drug group, while the expression of p62 protein was lower in the single-drug group. Fluorescence microscopy results showed that the number of EGFP-LC3 fluorescent aggregates in the combination group were more than that in the single-drug. Using amiodarone and glycyrrhetinic acid treated HepG2 cells, inhibition of auotophagy could decrease cell viability, increase apoptosis rate of cells; promoting autophagy would decrease the apoptosis rate and increase cells viability.

Conclusion: By increasing apoptosis of hepatocellular carcinoma HepG2 cells and autophagy level, and decreasing the cell activity, amiodarone combining with glycyrrhetinic acid treatment inducing autophagy a protective mechanism for cells.

Keywords: Amiodarone; Autophagy; Combined administration; Glycyrrhetinic acid; Liver cancer.