Enhancement of prostaglandin E2 production by epidermal growth factor requires the coordinate activation of cytosolic phospholipase A2 and cyclooxygenase 2 in human squamous carcinoma A431 cells.
We demonstrated the effect of epidermal growth factor (EGF) on the production of PGE2 in human squamous carcinoma A431 cells. The production of PGE2 was increased by stimulating the cells with EGF for 2 h and reached a maximum for 10 h. EGF was also found to augment the release of arachidonic acid (AA) following the increase in phospholipase A2 (PLA2) activity (1.7-fold). The induced PLA2 activity was diminished by 4-bromophenacyl bromide, but not by dithiothreitol, indicating that the EGF-induced release of AA was due to the increase in the activity of cytosolic PLA2 (cPLA2). On the other hand, cyclooxygenase (COX) activity was increased (1.6-fold) within 2 h after the EGF-treatment and the induced activity was inhibited by cycloheximide. In addition, Northern blot analysis showed that the level of COX-2 mRNA was increased by the EGF-treatment, whereas no COX-2 mRNA was detected in the untreated cells, indicating that the EGF-induced COX activity was resulted from the increase in the production of COX-2. These results suggest that EGF augments the production of PGE2 by increasing not only the activity of cPLA2 but also the production of COX-2 in A431 cells.
Sato
T
T
Department of Biochemistry, School of Pharmacy, Tokyo University of Pharmacy and Life Science, Japan. satotak@ps.toyaku.ac.jp
Nakajima
H
H
Fujio
K
K
Mori
Y
Y
D000098:Acetophenones / Q000494:pharmacology; D016718:Arachidonic Acid / Q000378:metabolism; D015152:Blotting, Northern; D002294:Carcinoma, Squamous Cell / Q000378:metabolism*; D003001:Cloning, Molecular; D003513:Cycloheximide / Q000494:pharmacology; D051546:Cyclooxygenase 2; D018076:DNA, Complementary / Q000235:genetics; D015232:Dinoprostone / Q000096:biosynthesis*; D004229:Dithiothreitol / Q000494:pharmacology; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000098:Acetophenones; D018076:DNA, Complementary; D004791:Enzyme Inhibitors; D007527:Isoenzymes; D008565:Membrane Proteins; D012333:RNA, Messenger; D016718:Arachidonic Acid; D004815:Epidermal Growth Factor; D003513:Cycloheximide; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011451:Prostaglandin-Endoperoxide Synthases; D010741:Phospholipases A; D054467:Phospholipases A2; D015232:Dinoprostone; C006821:4-bromophenacyl bromide; D004229:Dithiothreitol