Pertussis toxin-sensitive secretory phospholipase A2 expression and motility in activated primary human keratinocytes.
Secretory phospholipase A2 and cycloxygenase-2 are coexpressed in activated primary keratinocytes. These proteins are known to be functionally linked, mediating proliferation of human keratinocytes during epidermal wound repair. Primary human keratinocytes grown at low densities (15-30%; nonconfluent) produce high levels of prostaglandin E2 important for proliferation and are a good model for studying activated keratinocytes after injury. In this study, we used this model to assess the role of secretory phospholipase A2 and cycloxygenase-2 in keratinocyte motility. Initial work showed 24 h pretreatment with 20 ng pertussis toxin per ml, an inhibitor of the inhibitory G-protein, decreased prostaglandin E2 production and both secretory phospholipase A2 and cycloxygenase-2 protein expression. This suggested that inhibitory G-protein may be involved in mediating expression of these proteins. Pertussis toxin also caused changes in cell morphology, actin organization, and keratinocyte motility. Pretreatment with 5 microm 12-epi-scalaradial, a secretory phospholipase A2 inhibitor, caused similar changes in cell motility and actin organization; however, the specific cycloxygenase-2 inhibitor, SC-58236 (20 nm) was much less effective. These results suggested that secretory phospholipase A2 plays a part in keratinocyte motility that is independent of its functional linkage to cycloxygenase-2 and prostaglandin E2 biosynthesis.
Rys-Sikora
Krystyna E
KE
Department of Dermatology, University of Rochester Medical Center, Rochester, NY 14642, USA.
Pentland
Alice P
AP
Konger
Raymond L
RL
D016718:Arachidonic Acid / Q000378:metabolism; D002465:Cell Movement; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D015603:Keratinocytes / Q000201:enzymology* / Q000502:physiology; D037342:Pertussis Toxin / Q000494:pharmacology; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000502:physiology*; D054467:Phospholipases A2; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism; D011453:Prostaglandins / Q000096:biosynthesis
D004791:Enzyme Inhibitors; D011453:Prostaglandins; D016718:Arachidonic Acid; D011451:Prostaglandin-Endoperoxide Synthases; D037342:Pertussis Toxin; D010741:Phospholipases A; D054467:Phospholipases A2