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  • Title: Alteration of transcription factor mRNAs during the isolation and culture of rat hepatocytes suggests the activation of a proliferative mode underlies their de-differentiation.
    Author: Padgham CR, Boyle CC, Wang XJ, Raleigh SM, Wright MC, Paine AJ.
    Journal: Biochem Biophys Res Commun; 1993 Dec 15; 197(2):599-605. PubMed ID: 8267596.
    Abstract:
    The commonly observed loss of liver specific phenotype regularly described in rat hepatocyte culture is typified by the loss of total cytochrome P450 (CYP) content and the altered abundance of CYP mRNAs. The current work shows that these changes are preceded by the induction of the mRNA encoding the transcription factor c-jun during the hepatocyte isolation procedure. Then as the hepatocytes attach to the substratum the induced expression of c-jun subsides and two patterns of CYP mRNA loss are observed. The mRNAs encoding CYPs 2C11, 2C13, 2E1, 3A1, 3A2 and 4A1 continuously decline while CYP 1A2, 2A1/2 and 2B1/2 mRNAs are temporarily stabilised for 2 to 2.5 hours at a reduced level before declining further. The loss of CYP1A2 and 2B1/2 mRNAs parallels the loss of the mRNAs encoding the liver specific transcription factors C/EBP alpha and HNF-1. The early and rapid increase in c-jun mRNA followed by a decline in C/EBP alpha mRNA are characteristic of the changes in the expression of these transcription factor mRNAs following the stimulation of hepatocyte proliferation after partial hepatectomy. The finding that the rate of loss of total P450 following partial hepatectomy parallels that in rat hepatocyte culture suggests that the commonly employed hepatocyte isolation procedure "primes" the normally quiescent hepatocytes to enter the cell cycle and de-differentiate especially as both systems lose the major constitutively expressed CYP2C11 isozyme.
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