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Transformation of human liver L-O2 cells mediated by stable HBx transfection.
- Source :
-
Acta pharmacologica Sinica [Acta Pharmacol Sin] 2009 Aug; Vol. 30 (8), pp. 1153-61. Date of Electronic Publication: 2009 Jul 06. - Publication Year :
- 2009
-
Abstract
- Aim: To explore the mechanism of hepatocarcinogenesis associated with the hepatitis B virus X protein (HBx), we investigated the role of HBx in transformation using human liver L-O2 cells stably transfected with HBx as a model.<br />Methods: Plasmids encoding HBx were stably transfected into immortalized human liver L-O2 cells and rodent fibroblast NIH/3T3 cells. The expression of alfa-fetoprotein (AFP), c-Myc, HBx, and survivin in the engineered cells was examined by Western blotting. The malignant phenotype of the cells was demonstrated by anchorage-independent colony formation and tumor formation in nude mice. RNA interference assays, Western blotting, luciferase reporter gene assays and flow cytometry analysis were performed. The number of centrosomes in the L-O2-X cells was determined by gamma-tubulin immunostaining. The effect of HBx on the transcriptional activity of human telomerase reverse transcriptase (hTERT) and hTERT activity in L-O2-X cells and/or 3T3-X cells was detected by the luciferase reporter gene assay and telomerase repeat amplification protocol (TRAP).<br />Results: Stable HBx transfection resulted in a malignant phenotype in the engineered cells in vivo and in vitro. Meanwhile, HBx was able to increase the transcription of the NF-kappaB, AP-1, and survivin genes and to upregulate the expression levels of c-Myc and survivin. Abnormal centrosome duplication and activated hTERT were responsible for the transformation.<br />Conclusion: Stable HBx transfection leads to genomic instability of host cells, which is responsible for hepatocarcinogenesis; meanwhile, transactivation by the HBx protein contributes to the development of hepatocellular carcinoma (HCC). The L-O2-X cell line is an ideal model for investigating the mechanism of HBx-mediated transformation.
- Subjects :
- Animals
Cell Line
Cell Proliferation
Centrosome pathology
Gene Expression Regulation, Neoplastic
Humans
Inhibitor of Apoptosis Proteins
Mice
Mice, Nude
Microtubule-Associated Proteins genetics
Microtubule-Associated Proteins metabolism
NF-kappa B genetics
NF-kappa B metabolism
NIH 3T3 Cells
Survivin
Telomerase metabolism
Trans-Activators metabolism
Transcription Factor AP-1 genetics
Transcription Factor AP-1 metabolism
Viral Regulatory and Accessory Proteins
Carcinoma, Hepatocellular genetics
Cell Transformation, Neoplastic
Liver cytology
Liver Neoplasms genetics
Trans-Activators genetics
Transfection
Subjects
Details
- Language :
- English
- ISSN :
- 1745-7254
- Volume :
- 30
- Issue :
- 8
- Database :
- MEDLINE
- Journal :
- Acta pharmacologica Sinica
- Publication Type :
- Academic Journal
- Accession number :
- 19578387
- Full Text :
- https://doi.org/10.1038/aps.2009.99