Growth hormone signaling leading to CYP2C12 gene expression in rat hepatocytes involves phospholipase A2.
The expression of CYP2C12 is liver-specific and regulated at the transcriptional level by growth hormone (GH). In attempts to elucidate the nature of signaling molecules mediating the GH regulation of this gene in rat hepatocytes, a role for phospholipase A2 (PLA2) as a transducer of GH-induced levels of P4502C12 mRNA was investigated. GH was shown to induce tyrosyl-phosphorylation of p42 and p44 microtubule-associated protein (MAP) kinases and to reduce the electrophoretic mobility of a 100-kDa protein, immunologically related to cPLA2. These events were observed in parallel with GH-stimulated release of [3H]arachidonic acid ([3H]AA) from cellular phospholipids of rat hepatocytes labeled with [3H]AA. These rapid effects of GH action, as well as the GH-induced expression of CYP2C12, were inhibited in cells treated with the tyrosine kinase inhibitor herbimycin A. Similarly, when the GH-induced liberation of [3H]AA was blocked by the PLA2 inhibitor mepacrine or the Ca2+ channel blocker verapamil, GH-induced accumulation of P4502C12 mRNA was absent. These results suggest a correlation between PLA2 activity and GH regulation of the CYP2C12 gene. The inhibitory effect of mepacrine on GH induction of P4502C12 mRNA was reversed by AA addition, further supporting a role for eicosanoids in the regulation of CYP2C12. Finally, inhibitors of P450-mediated AA metabolism, SKF-525A and ketoconazole as well as eicosatetraynoic acid, blocked the GH-mediated induction of P4502C12 mRNA, whereas more specific inhibitors of cyclooxygenase or lipoxygenase metabolism did not. Based on these results, we suggest that GH signaling in rat hepatocytes, leading to increased expression of CYP2C12, involves PLA2 activation and subsequent P450-catalyzed formation of an active AA metabolite.
Tollet
P
P
Department of Medical Nutrition, Karolinska Institute, Huddinge University Hospital, Sweden.
Hamberg
M
M
Gustafsson
J A
JA
Mode
A
A
D015117:5,8,11,14-Eicosatetraynoic Acid / Q000494:pharmacology; D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D001189:Aryl Hydrocarbon Hydroxylases; D016227:Benzoquinones; D000001:Calcimycin / Q000494:pharmacology; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000096:biosynthesis* / Q000235:genetics; D004075:Diglycerides / Q000096:biosynthesis; D004789:Enzyme Activation; D005786:Gene Expression Regulation; D013006:Growth Hormone / Q000494:pharmacology*; D007334:Insulin-Like Growth Factor I / Q000096:biosynthesis / Q000235:genetics; D007654:Ketoconazole / Q000494:pharmacology; D047029:Lactams, Macrocyclic; D008099:Liver / Q000166:cytology / Q000187:drug effects*; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D010743:Phospholipids / Q000378:metabolism; D010766:Phosphorylation; D011335:Proadifen / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D011796:Quinacrine / Q000494:pharmacology; D011809:Quinones / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D017828:Rifabutin / Q000031:analogs & derivatives; D015398:Signal Transduction / Q000502:physiology*; D013250:Steroid Hydroxylases / Q000096:biosynthesis* / Q000235:genetics; D014700:Verapamil / Q000494:pharmacology
D016227:Benzoquinones; D004075:Diglycerides; D047029:Lactams, Macrocyclic; D010743:Phospholipids; D011809:Quinones; D012333:RNA, Messenger; D015117:5,8,11,14-Eicosatetraynoic Acid; D017828:Rifabutin; D016718:Arachidonic Acid; D000001:Calcimycin; D007334:Insulin-Like Growth Factor I; C020754:herbimycin; D013006:Growth Hormone; D003577:Cytochrome P-450 Enzyme System; D011335:Proadifen; D014700:Verapamil; D013250:Steroid Hydroxylases; D001189:Aryl Hydrocarbon Hydroxylases; C034694:steroid 15-beta-hydroxylase; D011505:Protein-Tyrosine Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D010741:Phospholipases A; D054467:Phospholipases A2; D011796:Quinacrine; D007654:Ketoconazole