PubMed · 14222571
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G W ARTHUR, J O STEWART. 1964. BILIARY CYSTS.. https://doi.org/10.1002/bjs.1800510907
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Biliary atresia(BA), the most common cause of obstructive jaundice in infancy, has been considered to be a result of progressive destruction of the bile ducts through a necroinflammatory process. Many immunohistochemical studies of BA remnant have been done, but it has not been shown that biliary epithelial cells (BECs) can be cultured from BA remnant. For this study, we obtained bile duct remnants from three patients with biliary atresia (one male, two females) who received Kasai's operation from 2002 to 2003. The successive cultivation rate of BECs from explants was 100% (3/3 patients). Culture of BECs on collagen gel was possible up to at least four passages. Under a phase-contrast microscope, primary and passed cultured cells on collagen gel showed a cobblestone-like spread in 2 weeks. The BECs had immunoreactivity to anti-human cytokeratin 7 antibody. In this study, we proved that BECs in the remnants of BA could be cultured, and defined the maturation of biliary epithelial cells of BA by immunocytochemistry with anti-human cytokeratin 7 antibody. In conclusion, BECs in BA remnant are "alive", and their proliferation activity can be maintained.
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AIM: 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. CONCLUSION: 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.