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This Article Full Text Full Text (PDF) All Versions of this Article: 295/2/G322    most recent 00597.2007v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Scheving, L. A. Articles by Russell, W. E. Search for Related Content PubMed PubMed Citation Articles by Scheving, L. A. Articles by Russell, W. E.

HORMONES AND SIGNALING

Cultured rat hepatocytes upregulate Akt and ERK in an ErbB-2-dependent manner

Departments of ¹Pediatrics, Division of Endocrinology and ²Cell and Developmental Biology, the ³Digestive Disease Research Center, the ⁴Vanderbilt Diabetes Center, and the ⁵Vanderbilt Ingram Cancer Center, Vanderbilt University Medical Center, Nashville, Tennessee

Submitted 21 December 2007 ; accepted in final form 2 June 2008

Epidermal growth factor (EGF) stimulates freshly plated adult hepatocytes to synthesize DNA, but only after they pass through a lag phase of 40 h following EGF exposure. The longer the cells are maintained, they become more responsive to EGF and the lag phase shortens. Maximal EGF-mediated stimulation of DNA synthesis requires the induction of ErbB2, which is not normally expressed in adult hepatocytes. We used immunological methods to demonstrate increased expression during culture of two gene families required for EGF to stimulate hepatocyte DNA synthesis: Akt and ERK 1/2. Both families showed hyperexpression in culture particularly when cells were exposed to insulin and EGF. Unlike CDK-2 and cyclin D1, integral mediators of the G1/S phase transition, ERK 1/2 and Akt appeared in the absence of EGF, particularly when insulin was present. This hyperexpression, which high concentrations of dexamethasone reversed, increased basal and growth factor-stimulated phosphorylation of Akt and ERK 1/2. Pharmacological blockade of phosphatidylinositol kinase suppressed the Akt increase whereas pharmacological blockade or small interfering RNA downregulation of ErbB2 inhibited both Akt and ERK 1/2 expression. All three Akt isoforms contributed to the increase in total Akt. EGF but not insulin specifically upregulated Akt 2 and 3. Since Akt and ERK 1/2 are also hyperexpressed in poorly differentiated hepatomas, their dysregulation in cancer may involve transcriptional mechanisms normally operative in cultured hepatocytes. We hypothesize that the induction and activation of ErbB2 increases the expression of these kinases, enhancing the responsiveness of hepatocytes to EGF as they adapt to culture.

cell culture; liver; signaling; EGF; insulin