Hepatitis B virus X gene induces human telomerase reverse transcriptase mRNA expression in cultured normal human cholangiocytes.
OBJECTIVE
To study the transcriptional regulation of human telomerase reverse transcriptase (hTERT) mRNA in normal human cholangiocytes (HBECs) after hepatitis B virus X (HBx) gene transfection and to elucidate the possible mechanism of HBV infection underlying cholangiocarcinoma.


METHODS
HBECs were cultured in vitro and co-transfected with a eukaryotic expression vector containing the HBx coding region and a cloning vector containing coding sequences of enhanced green fluorescent protein (EGFP) using lipid-mediated gene transfer. The transfection efficiency was determined by the expression of EGFP. The expressions of hTERT mRNA and HBx protein in HBECs were detected by RT-PCR and immunocytochemical stain, respectively.


RESULTS
The transfection efficiencies were about 15% for both HBx gene expression plasmid and empty vector. No hTERT mRNA was expressed in HBECs when transfected with OPTI-MEM medium and empty vector, but a dramatic increase was observed for hTERT mRNA expression in HBECs when transfected with HBx expression vector. HBx protein was only expressed in HBECs when transfected with HBx expression vector.


CONCLUSIONS
HBx transfection can activate the transcriptional expression of hTERT mRNA. Cis-activation of hTERT mRNA by HBx gene is the primary mechanism underlying the proliferation, differentiation and tumorigenesis of biliary epithelia.
Zou
Sheng-Quan
SQ
Department of Surgery, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China. sqzou@tjh.tjmu.edu.cn
Qu
Zhen-Liang
ZL
Li
Zhan-Fei
ZF
Wang
Xin
X
D001652:Bile Ducts / Q000166:cytology / Q000378:metabolism*; D002478:Cells, Cultured; D004268:DNA-Binding Proteins; D006801:Humans; D012333:RNA, Messenger / Q000378:metabolism*; D019098:Telomerase / Q000235:genetics*; D015534:Trans-Activators / Q000235:genetics / Q000502:physiology*; D014162:Transfection; D054334:Viral Regulatory and Accessory Proteins
D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein; D019098:Telomerase