Chronic-alcohol exposure alters IGF1 signaling in H9c2 cells via changes in PKC delta.
Previously, we have demonstrated that chronic-alcohol exposure alters insulin-like growth factor 1 (IGF1) signaling in adult rat heart cells. This report examines the effects of alcohol in vitro on the expression of protein kinase C (PKC) alpha, delta, and epsilon using the embryonic heart cell line, H9c2, and how this may be linked to changes in IGF1 signal transduction. Western blot analyses of H9c2 protein preparations demonstrate that there are significant increases in the total protein levels of PKC delta and epsilon after 4 days exposure to alcohol, and similar increases were found after 2 and 6 days exposure. In addition, there was a significant increase in PKC delta and epsilon in the membranal fractions and a decrease in the cytosolic fractions. No change was found in the expression or activity levels for PKC alpha. Chronic-alcohol exposure (100 mM, 4 days) increased the basal tyrosine kinase activity of the IGF1 receptor (IGF1R), and altered its rate of activation. Chronic-alcohol exposure also reduced the rate of Erk1/Erk2 activation by IGF1. Chronic alcohol blocked the proliferative effects of IGF1 on cell growth and reduced cell viability both in the presence and absence of IGF1, and this alcohol-induced reduction in cell viability was blocked using siRNA to inhibit PKC delta. In addition, a reduction in the amount of myosin light chain 2 was found in the alcohol-exposed cells. In conclusion, chronic alcohol alters PKC delta and epsilon expression and activity, and suppresses the IGF1 signaling pathway in embryonic heart cell culture. Blockage of PKC delta expression using siRNA inhibits the suppressive effects of alcohol on cell viability.
Ila
Richard
R
Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, PA 19107, USA.
Solem
Michele
M
D000199:Actins / Q000187:drug effects; D000818:Animals; D024722:Cardiac Myosins / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D000431:Ethanol / Q000494:pharmacology*; D048049:Extracellular Signal-Regulated MAP Kinases / Q000187:drug effects; D007334:Insulin-Like Growth Factor I / Q000187:drug effects*; D032383:Myocytes, Cardiac / Q000187:drug effects*; D018994:Myosin Light Chains / Q000187:drug effects; D051745:Protein Kinase C-delta / Q000187:drug effects*; D051744:Protein Kinase C-epsilon / Q000187:drug effects*; D034741:RNA, Small Interfering; D051381:Rats; D017526:Receptor, IGF Type 1 / Q000187:drug effects; D019209:Troponin C / Q000187:drug effects
D000199:Actins; D018994:Myosin Light Chains; D034741:RNA, Small Interfering; D019209:Troponin C; C107184:myosin light chain 2; D000431:Ethanol; D007334:Insulin-Like Growth Factor I; D017526:Receptor, IGF Type 1; D051745:Protein Kinase C-delta; D051744:Protein Kinase C-epsilon; D048049:Extracellular Signal-Regulated MAP Kinases; D024722:Cardiac Myosins