Increased expression of WAF1 in intrahepatic bile ducts in primary biliary cirrhosis relates to apoptosis

Abstract

Background/Aims : In primary biliary cirrhosis (PBC), the intrahepatic small bile ducts are selectively damaged by immune attacks, followed by progressive loss mainly due to apoptosis. Compared to the intercellular signaling such as the CD95/CD95 ligand interaction, little is known about alterations in intracellular cell cycle regulatory proteins and genotoxic damage in this apoptotic process. WAF1 is a potent and reversible inhibitor of cell cycle progression at both the G1 and G2 checkpoint and upregulated WAF1 induces irreversible G1 arrest and apoptosis. Transcriptional activation of the WAF1 gene is induced by the upregulated p53 in response to DNA damage. In this study, the cell cycle regulatory process of apoptosis in PBC was examined with respect to expression of WAF1. Methods : Immunostaining for WAF1 and p53 was performed using 11 liver sections of PBC and 26 control livers. In addition, Ki67, apoptosis (TUNEL-positive), and human telomerase RNA (hTR) were also detected. Results : WAF1 was expressed in the nuclei of several epithelial cells in most damaged bile ducts in PBC but infrequently or rarely in controls. Some of these cells were also positive for p53, while the remainder were not. Ki67 immunostaining and TUNEL disclosed that the bile ducts in PBC showed increased cell division as well as enhanced apoptosis. Immunostaining of Ki67 and TUNEL staining showed that WAF1-positive cells were not proliferating, while some WAF1-positive cells were undergoing apoptosis. Moreover, the bile ducts lacked hTR expression, implying progressive shortening of telomeres during increased cell divisions. Conclusions : It seems possible that in PBC, expression of WAF1 on biliary epithelial cells relates to the apoptosis. p53 may be involved in this upregulation. This may be due to physiological upregulation of WAF1 and p53 in response to genotoxic damage such as oxidative stress associated with cholangitis, suggesting other processes than CD95/CD95 ligand interaction in biliary epithelial apoptosis in PBC.