Characterization of liver function in transdifferentiated hepatocytes.
We previously demonstrated that dexamethasone (Dex) induces the transdifferentiation (or conversion) of the pancreatic progenitor cell line AR42J-B13 (B13) to hepatocytes based on the expression of liver proteins. We have extended our original observations to determine: (1) the effects of Dex on pancreatic gene expression; (2) the time course of expression of liver enriched transcription factors during conversion from pancreatic to hepatic phenotype; (3) the functional potential of transdifferentiated hepatocytes; (4) the proliferative capacity of transdifferentiated hepatocytes; and (5) whether ectopic expression of transcription factors can induce the hepatic phenotype in pancreatic B13 cells. The results were as follows. The B13 cell markers amylase, synaptophysin, and neurofilament were lost in transdifferentiated hepatocytes compared to control cells and the liver enriched transcription factors C/EBPbeta and C/EBPalpha were induced first, followed by HNF4alpha and then RXRalpha. Using RT-PCR analysis and immunolocalisation studies, we detected hepatic markers (e.g., apolipoprotein B) in Dex-treated cells. In transdifferentiated hepatocytes albumin was secreted, insulin stimulated lipid deposition and ciprofibrate enhanced the expression of catalase. Proliferation of transdifferentiated hepatocytes is promoted in the presence of HGF and NEAA as indicated by the co-expression of the cell cycle markers cyclin D and phosphohistone H3 with liver proteins. Lastly, ectopic expression of C/EBPalpha or C/EBPbeta in AR42J-B13 cells was sufficient to induce transdifferentiation, based on nuclear localization of HNF4alpha and induction of UDP-glucuronosyltransferase expression. These results indicate that the B13 progenitor cell model is suitable for studying liver function and for understanding the molecular and cellular events that occur during transdifferentiation.
Burke
Zoë D
ZD
Centre for Regenerative Medicine, Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath, United Kingdom.
Shen
Chia-Ning
CN
Ralphs
Kate L
KL
Tosh
David
D
D000418:Albumins / Q000378:metabolism; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D015415:Biomarkers / Q000378:metabolism; D022762:CCAAT-Enhancer-Binding Proteins / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D049109:Cell Proliferation; D003907:Dexamethasone / Q000494:pharmacology; D017228:Hepatocyte Growth Factor / Q000494:pharmacology; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D006801:Humans; D008055:Lipids / Q000737:chemistry; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D010179:Pancreas / Q000166:cytology*; D010641:Phenotype; D051381:Rats; D013234:Stem Cells; D014157:Transcription Factors / Q000378:metabolism; D015262:Xenobiotics / Q000494:pharmacology
D000418:Albumins; D000893:Anti-Inflammatory Agents; D015415:Biomarkers; D022762:CCAAT-Enhancer-Binding Proteins; D008055:Lipids; D014157:Transcription Factors; D015262:Xenobiotics; D017228:Hepatocyte Growth Factor; D003907:Dexamethasone