Differential regulation of hepatocyte dna synthesis by camp in vitro and in vivo . Westwick, Jk, J Fleckenstein, M Yin, Sq Yang, Ca Bradham, Da Brenner, Am Diehl. Department of Pharmacology and Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, NC 27599-7365, Department of Medicine, Johns Hopkins University, Baltimore, MD and Departments of Medicine and Biochemistry and Biophysics, University of North Carolina, Chapel Hill, NC 27599-7080
APStracts 3:0099G, 1996.
Cyclic AMP prevents epidermal growth factor (EGF)-induced DNA synthesis in many types of cultured cells including hepatocytes, but its effects on cellular proliferation in vivo are unknown. This study: 1) compares the effects of supplemental cAMP on hepatocyte proliferation induced in vivo by 70% partial hepatectomy (PH) and in vitro by epidermal growth factor (EGF), and 2) determines the effects of cAMP on AP-1, a family of growth-regulatory transcription factors, and the kinase cascades which normally activate AP-1. Although injection of dibutyryl cAMP (30 mg/kg i.p.) at the time of PH increased liver cAMP concentrations at least 100 fold for several hours, it did not inhibit hepatic incorporation of [3H] thymidine or PCNA expression 24 h after PH. cAMP treatment led to a complete inhibition of ERK activity and transiently reduced Jun N-terminal Kinase (JNK) activity after PH but did not decrease the expression of c-jun mRNA or protein. Consistent with the known cAMP stimulation of junB in cultured cells, cAMP treatment increased junB mRNA, protein and DNA binding activity post-PH. Surprisingly, cAMP treatment enhanced Raf kinase activity after PH in rats. In primary hepatocyte cultures, supplemental cAMP inhibited JNK and ERK activity, total AP -1 and c-Jun transcriptional activities, and DNA synthesis. Thus, elevated cAMP inhibited ERK and JNK activity in culture and in vivo, and inhibited hepatocyte proliferation in culture but not in vivo. This suggests that in vivo mechanisms compensate for cAMP inhibition of certain growth-related signaling cascades and emphasizes potential risks of extrapolating from simple cell culture systems to explain physiology in intact animals.

Received 3 January 1996; accepted in final form 19 April 1996.
APS Manuscript Number G7-6.
Article publication pending Am. J. Physiol. (Gastrointest. Liver
Physiology).
ISSN 1080-4757 Copyright 1996 The American Physiological Society.
Published in APStracts on 8 May 96