Functional expression of human heme oxygenase-1 (HO-1) driven by HO-1 promoter in vitro and in vivo.
We developed a retrovirus-mediated human heme oxygenase-1 (HO-1) gene expression system and assessed the impact of heme on the inducibility of the HO-1 gene in rat lung microvessel (RLMV) endothelial cells and in newborn Sprague-Dawley (SD) rats. Overexpression of the HO-1 gene driven by HO-1 promoter (HOP) resulted in an increase in HO-1 protein and HO activity by 4.8- and 1.3-fold, respectively, compared to the viral LTR promoter. The increased HO-1 gene expression was associated with the enhancement of CO production. In cells transduced by HOP-driven HO-1 gene, there was a decrease in basal cyclooxygenase (COX) activity as measured by PGE(2). The degree of HO-1 expression and, consequently, the levels of cellular heme were directly related to COX activity. Supplementation with heme markedly increased PGE(2) and cGMP synthesis. In all (6/6) of newborn SD rats injected with retrovirus LSN-HOP-HO-1, both HO-1 and neo(r) transcripts were expressed in tissues. We hypothesize that degree of HO-1 gene expression resulted in a differential rate of cellular heme-dependent enzyme gene expression, which may play a vital role in maintaining cellular homeostasis.
Quan
Shuo
S
Department of Pharmacology, New York Medical College, Valhalla, New York 10595, USA.
Yang
Liming
L
Shenouda
Sylvia
S
Jiang
Houli
H
Balazy
Michael
M
Schwartzman
Michal L
ML
Shibahara
Ichiyo
I
Shinohara
Kousei
K
Abraham
Nader G
NG
D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western; D002248:Carbon Monoxide / Q000378:metabolism; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000378:metabolism; D015232:Dinoprostone / Q000378:metabolism; D004730:Endothelium, Vascular / Q000201:enzymology; D015870:Gene Expression; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D005822:Genetic Vectors; D006418:Heme / Q000378:metabolism / Q000494:pharmacology; D006419:Heme Oxygenase (Decyclizing) / Q000235:genetics* / Q000378:metabolism; D051547:Heme Oxygenase-1; D006801:Humans; D008168:Lung / Q000098:blood supply; D008565:Membrane Proteins; D051379:Mice; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger; D051381:Rats; D017207:Rats, Sprague-Dawley; D012190:Retroviridae / Q000235:genetics
D008565:Membrane Proteins; D012333:RNA, Messenger; D006418:Heme; D002248:Carbon Monoxide; C496541:HMOX1 protein, human; D006419:Heme Oxygenase (Decyclizing); D051547:Heme Oxygenase-1; C496542:Hmox1 protein, mouse; D006152:Cyclic GMP; D015232:Dinoprostone