A Catalogue of Gene Expression Difference in Biliary Cirrhosis due to Biliary Atresia Using Differential Expressed Sequence Tags(EST) Screening

ABSTRACT

PURPOSE: Extrahepatic biliary atresia is the most common indication for liver transplantation in children, but the etiology of this disorders remains unknown. It would be very signficant to identify genes that are specifically expressed in pathologic liver tissue of biliary atresia and analyze the pattern of expression in those genes. METHODS: We made dot blot panels consisting of 1,730 different EST (expressed sequence tags) clones which were isolated from human hair dermal papilla cell cDNA library. Liver tissues were taken from a recipient with biliary atresia and a normal donor during living-related liver transplantation. Total RNA was extracted from each sample and reversely transcribed to make cDNA. Then radiolabelled cDNA probe pools were made by random primed DNA labeling method and used for screening differentially expressed genes using EST dot blot panel. RESULTS: Among the total of 1,730 EST clones, 26 cDNA clones were overexpressed in biliary cirrhosis. They revealed homology to genes encoding bcl-w, laminin binding protein, hepatocyte growth factor-regulated tyrosine kinase substrate (HRS), thymosin beta-4, 10; transforming growth factor (TGF)-beta, tissue inhibitor of metalloproteinase (TIMP)-1, signal recognition particle (SRP)4, eukaryotic initiation factor (eIF)-2alpha kinase, lysyl oxidase, aldolase A, gamma-glutamylcystein synthetase, collagen type I alpha1, 2, collagen type III, fibronectin, osteonectin, insulin-like growth factor binding protein (IGFBP)-2, 3, and more. In addition, the expression of 2 clones showed that gastrula zinc finger protein and one novel gene were decreased in biliary atresia. CONCLUSOIN This study identified differentially expressed genes in biliary cirrhosis from progressive biliary atresia using differential EST screening technique.