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© 1989 Oxford University Press

research-article

Studies of phospholipase and cyclooxygenase activities in herpes simplex virus type 2-transformed rat cells

Catherine R.Krebs Heitman 1, Moseley Waite 2 and Louis S. Kucera

Departments of Microbiology and Immunology Winston-Salem, NC 27103, USA
1Present address: Cellular and Genetic Toxicology Branch, Natlonal Institute of Environmental Health Sciences Research Triangle Park, NC 27709, USA
2Biochemistry, Bowman Gray School of Medicine of Wake Forest University Winston-Salem, NC 27103, USA

In this report we describe results showing that both arachidonic acid (20:4) release and prostaglandin (PG) synthesis in herpes simplex virus type 2 (HSV-2)-transformed tumorderived rat fibrosarcoma (RFS) cells are inhibited by indomethacin (cyclooxygenase inhibitor). These data suggest that cyclooxygenase (prostaglandin endoperoxide synthetase, PES) and phospholipase are coupled in their regulation in RFS cells. Data obtained using a cell-free assay to measure directly cyclooxygenase specific activity in the absence of phospholipase activity indicate that constitutive cyclooxygenase activity is very low in non-transformed REF and is not induced by 12-O-tetradecanoylphorbol-13-acetate (TPA). In contrast, after an initial degradation of constitutive enzyme, cyclooxygenase is synthesized de novo following TPA treatment of HSV-2-transformed RFS clonal cells. At least a 5-fold induction of cyclooxygenase occurs by 6 h post-treatment of HSV-2-transformed RFS clonal cells. This induction is inhibited by pretreatment of cells with cyclohximide (protein synthesis inhibitor) 1 h prior to TPA treatment. Results obtained following delayed addition of cycloheximide to cells after TPA treatment indicate that the newly synthesized cyclooxygenase has a half-life of ~5 min. In addition, HSV-2-transformed RFS clonal cells at low passage (< 10) show over a 3-fold greater increase in cyclooxygenase specific activity after TPA treatment compared to that in TPA-treated high passage (>20) cells, suggesting that induction of high levels of PG synthesis is transient in these cells. These data indicate that HSV-2 transformation of REF cells is associated with alteration of cyclooxygenase regulation.


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