Selective augmentation of prostacyclin production by combined prostacyclin synthase and cyclooxygenase-1 gene transfer.
BACKGROUND
We tested the hypothesis that combined cyclooxygenase-1 (COX-1) and prostacyclin synthase (PGIS) gene transfer selectively augments prostacyclin production without a concurrent overproduction of other prostanoids.


RESULTS
ECV304 cells were transfected with bicistronic pCOX-1/PGIS versus pCOX-1 or pPGIS, and prostanoids were analyzed. Contrary to the high prostaglandin E2 synthesis in pCOX-1 transfected cells, selective prostacyclin formation was noted with bicistronic plasmid transfection. Next, we determined the optimal ratio of Ad-COX-1 to Ad-PGIS by transfecting human umbilical vein endothelial cells with various titers of these 2 adenoviral constructs and determined the level of protein expression and prostanoid synthesis. Our results show that optimal ratios of adenoviral titers to achieve a large prostacyclin augmentation without overproduction of prostaglandin E2 or F2alpha were 50 to 100 plaque forming units (pfu) of Ad-COX-1 to 50 pfu of Ad-PGIS per cell. A higher Ad-PGIS to Ad-COX-1 ratio caused a paradoxical decline in prostacyclin synthesis.


CONCLUSIONS
Prostacyclin synthesis can be selectively augmented by cotransfecting endothelial cells with an optimal ratio of COX-1 to PGIS. Combined COX-1 and PGIS gene transfer has the potential for therapeutic augmentation of prostacyclin.
Shyue
S K
SK
Vascular Biology Program, Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan.
Tsai
M J
MJ
Liou
J Y
JY
Willerson
J T
JT
Wu
K K
KK
D015121:6-Ketoprostaglandin F1 alpha / Q000032:analysis / Q000096:biosynthesis; D000256:Adenoviridae / Q000235:genetics; D016718:Arachidonic Acid / Q000378:metabolism; D002460:Cell Line; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D051545:Cyclooxygenase 1; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D015232:Dinoprostone / Q000032:analysis / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D011464:Epoprostenol / Q000032:analysis / Q000096:biosynthesis*; D005231:Fatty Acids, Unsaturated / Q000032:analysis / Q000096:biosynthesis; D022761:Gene Transfer, Horizontal; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D006801:Humans; D019746:Intramolecular Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins; D010957:Plasmids / Q000235:genetics; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
D005231:Fatty Acids, Unsaturated; D007527:Isoenzymes; D008565:Membrane Proteins; D016718:Arachidonic Acid; C008033:12-hydroxy-5,8,10-heptadecatrienoic acid; D015121:6-Ketoprostaglandin F1 alpha; D003577:Cytochrome P-450 Enzyme System; D011464:Epoprostenol; D051545:Cyclooxygenase 1; C496511:PTGS1 protein, human; D011451:Prostaglandin-Endoperoxide Synthases; D019746:Intramolecular Oxidoreductases; C022461:prostacyclin synthetase; D015232:Dinoprostone