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http://dx.doi.org/10.5352/JLS.2016.26.7.764

Apoptotic Effect of Extract from Artemisia annua Linné by Akt/mTOR/GSK-3β Signal Pathway in Hep3B Human Hepatoma Cells  

Kim, Eun Ji (Department of Biological Science and Biotechnology, College of Life Science and Nano Technology, Hannam University)
Kim, Guen Tae (Department of Biological Science and Biotechnology, College of Life Science and Nano Technology, Hannam University)
Kim, Bo Min (Department of Biological Science and Biotechnology, College of Life Science and Nano Technology, Hannam University)
Lim, Eun Gyeong (Department of Biological Science and Biotechnology, College of Life Science and Nano Technology, Hannam University)
Ha, Sung Ho (Department of Chemical Engineering, College of Life Science and Nano Technology, Hannam University)
Kim, Sang-Yong (Department of Food Science & Bio Technology, Shinansan University)
Kim, Young Min (Department of Biological Science and Biotechnology, College of Life Science and Nano Technology, Hannam University)
Publication Information
Journal of Life Science / v.26, no.7, 2016 , pp. 764-771 More about this Journal
Abstract
Extracts from Artemisia annua Linné (AAE) have been known to possess various functions, including anti-bacterial, anti-virus, and anti-oxidant effects. However, the mechanism of those effects of AAE is not well-known. The aim of this study was to analyze the inhibitory effects of AAE on cell proliferation of the human hepatoma cell line (Hep3B) and to examine its effects on apoptosis. Activation by phosphorylation of Akt is cell proliferation through the phosphorylation of TSC2, mTOR, and GSK-3β. We suggested that AAE may exert cancer cell apoptosis through Akt/mTOR/GSK-3β signal pathways and mitochondria-mediated apoptotic proteins. For this, we examined the effects of extracts of AAE on cell proliferation according to treatment concentration. Treatment with AAE not only reduced cell viability, but also resulted in the induced release of lactate dehydrogenase (LDH). These results were determined with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and a lactate dehydrogenase (LDH) assay. Furthermore, we determined the effects of apoptosis through Hoechst 33342 staining, annexinⅤ-propidium iodide (PI) staining, 5,5′, 6,6′-tetrachloro-1,1′,3,3′-tetraethyl-imidacarbocyanine iodide (JC-1) staining, and Western blotting. Our study showed that the treatment of liver cancer cells with AAE resulted in the inhibition of Akt, TSC2, GSK-3β-phosphorylated, Bcl-2, and pro-caspase 3 and the activation of Bim, Bax, Bak, and cleaved PARP expressions. These results indicate that AAE induced apoptosis by means of a mitochondrial event through the regulate of Akt/mTOR/GSK-3β signaling pathways.
Keywords
Akt/mTOR/GSK-3β pathway; Bax-Bak; bim; Hep3B; mitochondria potential;
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