Bicyclol induces cell cycle arrest and autophagy in HepG2 human hepatocellular carcinoma cells through the PI3K-AKT and Ras-Raf-MEK-ERK pathwaysReport as inadecuate




Bicyclol induces cell cycle arrest and autophagy in HepG2 human hepatocellular carcinoma cells through the PI3K-AKT and Ras-Raf-MEK-ERK pathways - Download this document for free, or read online. Document in PDF available to download.

BMC Cancer

, 16:742

Cell and molecular biology

Abstract

BackgroundBicyclol, 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.

MethodsIn 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.

ResultsBicyclol 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.

ConclusionThese 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.

KeywordsBicyclol Cell cycle Autophagy AKT ERK HepG2 Abbreviations3-MA3-methyladenine

AKTv-akt murine thymoma viral oncogene homolog

CDKCyclin-dependent kinases

ERKExtracellular signal-regulated kinases

FITCFluorescein isothiocyanate

GFPGreen fluorescent protein

LC3Light chain 3

Mdm2Mouse double minute 2 homolog

MEKMitogen-extracellular signal-regulated kinase

mTORMammalian target of rapamycin

mTORC1mTOR complex 1

NF-kBNuclear factor kappa-light-chain-enhancer of activated B cells

PAGEPolyacrylamide gel electrophoresis

PBSPhosphate buffer saline

PIPropidium iodide

PI3KPhosphatidylinositide 3-kinases

RbRetinoblastoma protein

RFPRed fluorescent protein

siRNASmall interfering RNA

TSCTuberous sclerosis complex

Electronic supplementary materialThe online version of this article doi:10.1186-s12885-016-2767-2 contains supplementary material, which is available to authorized users.

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Author: Yu Wang - Hao Nie - Xin Zhao - Yong Qin - Xingguo Gong

Source: https://link.springer.com/



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