TY - JOUR T1 - PI3K Inhibitors Reverse the Suppressive Actions of Insulin on CYP2E1 Expression by Activating Stress-Response Pathways in Primary Rat Hepatocytes JF - Molecular Pharmacology JO - Mol Pharmacol SP - 1138 LP - 1146 DO - 10.1124/mol.59.5.1138 VL - 59 IS - 5 AU - Jaspreet S. Sidhu AU - Fei Liu AU - Sean M. Boyle AU - Curtis J. Omiecinski Y1 - 2001/05/01 UR - http://molpharm.aspetjournals.org/content/59/5/1138.abstract N2 - Insulin-associated signaling pathways are critical in the regulation of hepatic physiology. Recent inhibitor-based studies have implicated a mechanistic role for phosphatidylinositol 3′ kinase (PI3K) in the insulin-mediated suppression of CYP2E1 mRNA levels in hepatocytes. We investigated the dose dependence for this response and for the effects of insulin and extracellular matrix on PI3K signaling and CYP2E1 mRNA expression levels using a highly defined rat primary hepatocyte culture system. The PI3K inhibitors wortmannin and LY294002 stimulated stress-activated protein kinase/c-Jun NH2-terminal kinase (SAPK/JNK) and p38 mitogen-activated protein kinase (MAPK) phosphorylation in a rapid and concentration-dependent manner that paralleled the inhibition of protein kinase B (PKB) phosphorylation. Although PI3K inhibitors reversed the suppressive effects of insulin on CYP2E1 expression, these effects only occurred at concentrations well in excess of those required to achieve complete inhibition of PKB phosphorylation. These same concentrations produced cytotoxic responses as evidenced by perturbed cellular morphology and elevated release of lactate dehydrogenase. Wortmannin-mediated activation of the SAPK/JNK and p38 MAPK pathways also resulted in the mobilization of activator protein-1 complex to the nuclear compartment. We conclude that the suppression of CYP2E1 mRNA expression by insulin is not directly associated with PI3K-dependent pathway activation, but rather is linked to a cytotoxic response stemming from acute challenge with PI3K inhibitors. ER -