title: Bicyclol induces cell cycle arrest and autophagy in HepG2 human hepatocellular carcinoma cells through the PI3K/AKT and Ras/Raf/MEK/ERK pathways.
abstract: BACKGROUND
  Bicyclol, a novel synthetic antihepatitis drug, is widely known to protect against liver injury. However, few reports have focused on the possible effect of bicyclol on anti-proliferation and autophagy induction in cancer cells, particularly hepatocellular carcinoma cells.
  
  
  METHODS
  In this study, we investigated the antitumor efficacy of Bicyclol in HepG2 cells and the mechanism of cell growth inhibition. Cell proliferation was analyzed by MTT assay, and the cell cycle and apoptosis were assessed by flow cytometry. And we transfected the cells with the GFP-RFP-LC3 vector to detect the autophagy flux in the cells. Mechanisms of bicyclol-induced cell growth inhibition were probed by western blot analysis.
  
  
  RESULTS
  Bicyclol effectively inhibited HepG2 cell proliferation in a dose- and time-dependent manner. In addition, we found that bicyclol inhibited cell cycle progression at G1 phase and induced autophagy in HepG2 cells, which implied that the significant decrease in cell proliferation was mainly induced by autophagy and inhibition of cell proliferation. Furthermore, western blot showed that bicyclol inhibited phosphorylation of Akt and ERK, down-regulated the expressions of cyclin D1, cyclin E2, CDK2, CDK4, p-Rb and p-mTOR. Moreover, AKT or ERK knockdown by siRNA enhanced bicyclol-induced autophagy and inhibition of cell proliferation.
  
  
  CONCLUSIONS
  These results suggest that bicyclol has potent anti-proliferative activity against malignant human hepatoma cells via modulation of the PI3K/AKT pathway and the Ras/Raf/MEK/ERK pathway, and indicate that bicyclol is a potential liver cancer drug worthy of further research and development.
authors.affiliation: Institute of Biochemistry, College of Life Sciences, Zijingang campus, Zhejiang University, Room 345, Hangzhou, 310058, Zhejiang, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Institute of Biochemistry, College of Life Sciences, Zijingang campus, Zhejiang University, Room 345, Hangzhou, 310058, Zhejiang, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Nie
authors.affiliation: Institute of Biochemistry, College of Life Sciences, Zijingang campus, Zhejiang University, Room 345, Hangzhou, 310058, Zhejiang, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Institute of Biochemistry, College of Life Sciences, Zijingang campus, Zhejiang University, Room 345, Hangzhou, 310058, Zhejiang, China. happy_ququ@126.com.
authors.forename: Yong
authors.initials: Y
authors.lastname: Qin
authors.affiliation: Institute of Biochemistry, College of Life Sciences, Zijingang campus, Zhejiang University, Room 345, Hangzhou, 310058, Zhejiang, China. gongxg@zju.edu.cn.
authors.forename: Xingguo
authors.initials: X
authors.lastname: Gong
keywords: AKT; Autophagy; Bicyclol; Cell cycle; ERK; HepG2