Abstract
Background and Aims: Gallbladder cancer (GBC) is the most common malignancy of the biliary tract. The prognosis for patients with GBC is poor as diagnosis is often at late untreatable stages of the disease. Chronic cholelithiasis has been linked to GBC. However, our understanding of the underlying mechanisms for the development of GBC and its association with gallstones (GS) is not clear. The aim of the present study was to establish and characterize a gallbladder cell line to study the molecular mechanisms of pathogenic relationship between gallstone formation and the development of gallbladder cancer. Methods: Primary cultures of gallbladder epithelial cells (GBECs) from prairie dogs were immortalized following previous hTERT method. Sub-cellular organelles were examined under electron microscope(EM) and immunohistochemical analysis was performed to examine the expression of biliary epithelial specific cytokeratines, vimentin and tumor supressor genes. RT-PCR analysis was performed to determine the expression of major Na+ transporter, Na+/H+ exchangers (NHEs) andMegalin. Results: Immortalized GBECs (GBECT) grew twelve passes and became confluent within 48 hrs. EM revealed distinct microvilli andmitochondria. GBECT coexpressed epithelial markers keratins and mesenchymal origin vimentin and showed strong p53 expression. GBECT retained absorbing property of gallbladder by expressing NHE1-3 isoforms. Conclusion: This is a novel gallbladder cell line from the prairie dog that retains gallbladder basic absorbing characteristic while acquires some tumorgenecity and may serve as an important model to study the underlying mechanisms of pathogenic role of GS formation on GBC development.
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