title: Regulation of expression of the human heme oxygenase-1 gene in transfected chick embryo liver cell cultures.
abstract: Induction of heme oxygenase (HO) has been proposed as a protective cellular mechanism against oxidative damage. In previous work (Tyrrell et al., Carcinogenesis [1993] 14, 761-765), portions of the 5' promoter region of the human HO-1 gene linked to the reporter gene chloramphenicol acetyl transferase (CAT), had been transiently expressed in HeLa cells. To extend the study of human HO gene expression into primary liver cells, these reporter gene fusion constructs, containing 121 or 1416 base pairs of the untranscribed 5'-upstream sequences of the human HO-1 gene, were used along with pSV beta-Gal plasmid to dually transfect primary cultures of chick embryo liver cells (CELC). The transfected cells were treated with selected metals, heme, phorbol ester, and chemical agents that produce oxidative stress (H2O2 or sodium arsenite). Reporter gene activities were measured 18-20 h later. Our major findings are: (1) these HO-CAT constructs were expressed in CELC; (2) unlike HeLa cells, the expression of CAT was detected in CELC without the need for the SV40 enhancer; (3) sodium arsenite and cobalt chloride induced the expression of the HO-CAT constructs whereas heme had no effect on or decreased CAT expression for all of the transfected constructs; (4) study of endogenous chick HO-1 gene expression in CELC showed that HO-1 responded to sodium arsenite treatment in a dose-dependent fashion, and the response was rapid and transient. We conclude that, in chick liver cell cultures, induction of the HO-1 gene by heme is fundamentally different from that produced by transition metals or sodium arsenite. Furthermore, the results suggest that expression of the HO-1 gene is highly conserved across species.
authors.affiliation: Department of Medicine, University of Massachusetts Medical Center, North Worcester 01655, USA.
authors.forename: T H
authors.initials: TH
authors.lastname: Lu
authors.forename: J A
authors.initials: JA
authors.lastname: Pepe
authors.forename: O S
authors.initials: OS
authors.lastname: Gildemeister
authors.forename: R M
authors.initials: RM
authors.lastname: Tyrrell
authors.forename: H L
authors.initials: HL
authors.lastname: Bonkovsky
mesh_terms: D000818:Animals; D018053:Arsenites; D002478:Cells, Cultured; D002642:Chick Embryo; D003035:Cobalt; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006418:Heme; D006419:Heme Oxygenase (Decyclizing) / Q000235:genetics*; D051547:Heme Oxygenase-1; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D008099:Liver / Q000196:embryology; D008565:Membrane Proteins; D012313:RNA / Q000302:isolation & purification; D017670:Sodium Compounds; D014162:Transfection
chemical_list: D018053:Arsenites; D008565:Membrane Proteins; D017670:Sodium Compounds; D003035:Cobalt; D006418:Heme; C017947:sodium arsenite; D012313:RNA; D007770:L-Lactate Dehydrogenase; C496541:HMOX1 protein, human; D006419:Heme Oxygenase (Decyclizing); D051547:Heme Oxygenase-1