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

WEI Qing-zhu, HE Li-li, WANG Xiao-yun. et al

Abstract

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, Glycyrrhetinic acid, Autophagy, Combined administration, Liver cancer 

 

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References


DING XX, ZHU QG, ZHANG SM, et al. Precision medicine for hepatocellular carcinoma; driver mutations and targeted therapy. Oncotarget,2017.8(33);55715-55730.

XIAO S. LIU Z, DENG R. et al. Aptamer-mediated gene therapy enhanced antitumor activity against human hepatocellular carcinoma in vitro and in vivo. J Control Release,2017,258(1): 130-145.

ISOMOTO S. KAWAKAMI A, OHTSURU A. et al. Antiarrhythmic amiodarone mediates apoptotic cell death of hepg2 hepatoblastoma cells through the mitochondrial pathway. Acta Med Nagasaki,2004 ,49( 1); 13-17.

LAN SH* WU SY, ZUCHINI R, et al. Autophagy suppresses tumorigenesis of hepatitis В virus-associated hepatocellular carcinoma through degradation of microRNA- 224. Hepatology,2014,59(2):505-517.

VASSALLO P. TROHMAN RG. Prescribing amiodarone. J AM A, 2007,298( 11): 1312-1322.

CAI Y, XU Y, CHEN HF, et al. Glycyrrhetinic acid mediated drug delivery carriers for hepatocellular carcinoma therapy. Mol Pharma.2016 ,13(3) ;699-709.

TANG ZH, LI T, CHANG LL, et al. Glycyrrhetinic acid triggers a protective autophagyby activation of extracellular regulated protein kinases in hepatocellular carcinoma cells. J Agric Food Chem,2014,62(49): 11910-11916.

EDINGER AL, THOMPSON CB. Death by design: apoptosis, necrosis and autophagy. Curr Opin Cell Biol, 2004,16(6):663-669.

GOLI-BENNOURA EE. ZOUAOUI O, NOUIRA S, et al. Cytotoxicity effects of amiodarone on cultured cells. Exp Toxicol Pathol,2012,64(5);425-430.

BAEHRECKE EH. Autophagy: dual roles in life and death? Nat Rev Mol Cell Biol,2005,6(6);505-510.

WANG Q, ZHU J, ZHANG K. et al. Induction of cytoprotective autophagy in PC-12 cells by cadmium. Biochem Biophys Res Commun,2013,438( 1): 186-192.

LI R. ZHOU P, GUO Y, et al. Tris (1, 3-dichloro-2- propyl) phosphate induces apoptosis and autophagy in SH- SY5Y cells: Involvement of ROS-mediated AMPK/mTOR/ ULK1 pathways. Food Chem Toxicol, 2017, 100 ( 1 ): 183- 196.

TANG ZH, ZHANG LL, LI T, et al. Glycyrrhetinic acid induces cytoprotective autophagy via the inositol-requiring enzyme la-c-Jun N-terminal kinase cascade in non-small cell lung cancer cells. ()ncotarget,2015 ,6(41):43911-43926.


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