효과로 AMPK 와 Akt 에 활성에도 관여하는 것으로 추측하고 있으며 추가 실험이 필요하다.
참 고 문 헌
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- ABSTRACT -
AMP-activated protein kinase-mTOR signaling pathway and hepatitis B virus replication
Jumi Kim
Department of Biomedical science The Graduate School, Ajou University
(Supervised by Associate Professor Kyongmin Kim)
Hepatitis B virus(HBV) is a small and enveloped DNA virus, belongs to family Hepadnaviridae. 350 million people worldwide are chronically infected. HBV causes acute
and chronic hepatitis B infections. Patients with chronic hepatitis B have a risk of developing cirrhosis and hepatocellular carcinoma(HCC). AMP-activated protein kinase(AMPK), an energy sensor, is a necessary protein to maintain energy balance. Even though hepatitis C virus(HCV) replication decreases AMPK activity and increases mammalian target of rapamycin(mTOR) activity, nothing has been known regarding HBV replication and AMPK activity. HBV X protein(HBx) had been reported to up-regulate mTOR activity and contribute to HCC development. In the present study, we examined whether HBV replication can regulate AMPK-mTOR signaling pathway or vice versa. Expression and
phosphorylation of AMPK were analyzed in the HepG2-derived stable HBV replicating cells, HepG2.2.15 and PEB 8 cells, and the Huh7-derived stable HBV replicating cells. We found that phosphorylation of AMPK were down-regulated in the HepG2.2.15 cells and PEB8 cells and the Huh7 HBV WT stable cells. We found that expression and phosphorylation of mTOR were up-regulated in HepG2.2.15 cells, PEB8 cells, and Huh7 HBV WT stable cells.
Because phosphorylation of AMPK was decreased, we expected that phosphorylation of regulatory subunit of mTOR (Raptor) and tuberous sclerosis 2 (TSC2) were decreased. But, phosphorylations of Raptor and TSC2 were increased in HepG2.2.15 cells. We found that phosphorylations of Akt was up-regulated in HepG2.2.15 cells. We found that expressions and phosphorylations of PP2A, S6K1 and 4E-BP1 were up-regulated in HepG2.2.15 cells.
And expressions and phosphorylations of PP2A and S6K1 were up-regulated in the Huh7 HBV WT stable cells. When expression and phosphorylation of AMPK were up-regulated via overexpression of AMPK alpha 2, HBV core particle, nucleic acid inside core particle, and HBV DNAs were reduced in HepG2 cells. When HepG2.2.15 cells were treated with 1mM metformin, AMPK activator, Akt, mTOR, PP2A and S6K1 phosphorylation were decreased. No significant changes in protein expressions. When HepG2.2.15 cells were treated with 10nM rapamycin, mTOR inhibitor, phosphorylations of mTOR and S6K1 were decreased. Surprisingly, phosphorylation of AMPK were increased and phosphorylation of Akt were decreased. We will further pursue which molecules are upstream or downstream of AMPK-mTOR signaling pathway.
Keyword : HBV, AMPK-mTOR signal, Akt, PP2A, S6K1, 4E-BP1