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KMID : 0613820160260070764
Journal of Life Science
2016 Volume.26 No. 7 p.764 ~ p.771
Apoptotic Effect of Extract from Artemisia annua Linne by Akt/mTOR/GSK-3¥â Signal Pathway in Hep3B Human Hepatoma Cells
Kim Eun-Ji

Kim Guen-Tae
Kim Bo-Min
Lim Eun-Gyeong
Ha Sung-Ho
Kim Sang-Yong
Kim Young-Min
Abstract
Extracts from Artemisia annua Linne (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-3¥â. We suggested that AAE may exert cancer cell apoptosis through Akt/mTOR/GSK-3¥â 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, annexin¥´-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-3¥â- 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-3¥â signaling pathways.
KEYWORD
Akt/mTOR/GSK-3¥â pathway, Bax-Bak, bim, Hep3B, mitochondria potential
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