Advertisement

Differential expression of ribosomal L31, Zis, gas-5 and mitochondrial mRNAs following oxidant induction of proliferative vascular smooth muscle cell phenotypes

      Abstract

      Treatment of cultured vascular smooth muscle cells (vSMCs) with benzo(a)pyrene (BaP), a prooxidant present in the particulate phase of tobacco smoke, induces highly proliferative (i.e. atherogenic) phenotypes. Critical early target genes in vSMCs have been identified, but patterns of gene expression following repeated cycles of carcinogen treatment in vivo have yet to be evaluated. In the present study, male Sprague–Dawley rats (175–200 g) were given weekly injections of BaP (10 mg/kg) for 8 weeks to induce atherogenic phenotypes. At the end of this atherogenic regimen, vSMCs were established in serial culture and monitored for patterns of proliferative activity and gene expression. vSMCs isolated from BaP-treated animals (hence forth referred to as BaP cells) exhibited constitutively increased growth rates, and marked enhancement of proliferation in response to serum mitogens. Differential display polymerase chain reaction (DD-PCR) and Northern blot analyses revealed that mRNAs for ribosomal protein L31 and Zis genes were suppressed, while gas-5 and mitochondrial mRNAs were overexpressed in BaP cells relative to control mRNA populations. In situ hybridization experiments in vascular tissue confirmed these alterations in vivo. This is the first report linking expression of these genes to proliferative dysregulation during the course of experimentally-induced atherogenesis.

      Keywords

      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'

      Subscribe:

      Subscribe to Atherosclerosis
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect

      References

      1. United States Department of Health and Human Services. The health consequences of smoking. Cardiovascular disease. 1991. A report of the surgeon general. Washington, DC: United States Printing Office.

        • Ramos K.S.
        Redox regulation of c-Ha-ras and osteopontin signaling in vascular smooth muscle cells: implications in chemical atherogenesis.
        Ann. Rev. Pharm. Toxicol. 1999; 39: 243-265
        • Ramos K.S.
        • Zhang Y.
        • Sadhu D.N.
        • Chapkin R.S.
        The induction of proliferative vascular smooth muscle cell phenotypes by benzo(a)pyrene is characterized by up-regulation of inositol phospholipid metabolism and c-Ha-ras gene expression.
        Arch. Biochem. Biophys. 1996; 332: 213-222
        • Lu K.P.
        • Ramos K.S.
        Identification of genes differentially expressed in vascular smooth muscle cells following benzo(a)pyrene challenge: implications for chemical atherogenesis.
        Biochem. Biophys. Res. Commun. 1998; 253: 828-833
        • Labrador M.
        • Vorces V.G.
        Transposable element-host interactions: regulation of insertion and excision.
        Ann. Rev. Genet. 1997; 31: 381-404
      2. Ramos KS, Cox LR. Aortic endothelial and smooth muscle cell cultures, In: Tyson CA, Frazier JM, editors. Methods in Toxicology, vol. 1A. In vitro biological systems, 1993. p. 159–68.

        • Ramos K.S.
        • Weber T.J.
        • Liau G.
        Altered protein secretion and extracellular matrix deposition is associated with the proliferative phenotype induced by allylamine in aortic smooth muscle cells.
        Biochem. J. 1993; 289: 57-63
        • Bradford M.M.
        A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
        Anal. Biochem. 1976; 72: 248-254
        • Altschul S.F.
        • Madden T.L.
        • Schaffer A.A.
        • Zhang J.
        • Zhang Z.
        • Miller W.
        • Lipman D.J.
        Gapped BLAST and PSI-BLAST: a new generation of protein database search programs.
        Nucl. Acids Res. 1997; 25: 3389-3402
        • Spencer T.E.
        • Gray C.A.
        • Joyce M.M.
        • Jenster G.
        • Wood C.G.
        • Bazer F.W.
        • Wiley A.A.
        • Bartol F.F.
        Discovery and characterization of genes expressed in the endometrial epithelium using the ovine uterine gland knockout model.
        Endocrinology. 1999; 140: 4070-4080
        • Sadhu D.N.
        • Ramos K.S.
        Cyclic AMP inhibits c-Ha-ras protooncogene expression and DNA synthesis in rat aortic smooth muscle cells.
        Experientia. 1993; 49: 567-570
        • Bonin L.R.
        • Madden K.
        • Shera K.
        • Ihle J.
        • Matthews C.
        • Aziz S.
        • Perez-Reyes N.
        • McDougall J.K.
        • Conroy S.C.
        Generation and characterization of human smooth muscle cell lines derived from atherosclerotic plaque.
        Arterioscl. Throm. Vasc. Biol. 1999; 19: 575-587
        • Ou X.
        • Ramos K.S.
        Proliferative response of quail aortic smooth muscle cells to benzo(a)pyrene: implications in PAH-induced atherogenesis.
        Toxicology. 1993; 74: 243-258
        • Tanaka T.
        • Kuwano Y.
        • Kuzumaki T.
        • Ishikawa K.
        • Ogata K.
        Nucleotide sequence of cloned cDNA specific for rat ribosomal protein L31.
        Eur. J. Biochem. 1987; 162: 45-48
        • Fabijanski S.
        • Pellegrini M.
        Identification of proteins at the peptidyl-tRNA binding site of rat liver ribosomes.
        Mol. Gen. Genet. 1981; 184: 5551-5556
        • Crawford D.R.
        • Wang Y.
        • Schools G.P.
        • Kochheiser J.
        • Davies K.J.A.
        Down-regulation of mammalian mitochondrial RNAs during oxidative stress.
        Free Radic. Biol. Med. 1997; 22: 551-559
        • Ou J.H.
        • Yen T.S.
        • Wang Y.F.
        • Kam W.K.
        • Rutter W.J.
        Cloning and characterization of a human ribosomal protein gene with enhanced expression in fetal and neoplastic cells.
        Nucl. Acids Res. 1987; 15: 8919-8934
        • Chester K.A.
        • Robson L.
        • Begent R.H.J.
        • Talbot I.C.
        • Pringle J.H.
        • Primrose L.
        • Macpherson A.J.S.
        • Boxer G.
        • Southall P.
        • Malcolm A.D.B.
        Identification of a human ribosomal protein mRNA with increased expression in colorectal tumours.
        Biochim. Biophys. Acta. 1989; 1009: 297-300
        • Shimbara N.
        • Sato C.
        • Takashina M.
        • Tanaka T.
        • Tanaka K.
        • Ichihara A.
        Down-regulation of ubiquitin gene expression during differentiation of human leukemia cells.
        FEBS. 1993; 322: 235-239
        • Karginova E.A.
        • Pentz E.S.
        • Kazakova I.G.
        • Norwood V.F.
        • Carey R.M.
        • Gomez R.A.
        Zis: a developmentally regulated gene expressed in juxtaglomerular cells.
        Am. J. Physiol. 1997; 273: F731-F738
        • Alejandro N.F.
        • Parrish A.R.
        • Bowes III, R.C.
        • Burghardt R.C.
        • Ramos K.S.
        Phenotypic profiles of cultured glomerular cells following repeated cycles of hydrocarbon injury.
        Kidney Int. 2000; 57: 1571-1580
        • Coccia E.M.
        • Cicala C.
        • Charlesworth A.
        • Ciccarelli C.
        • Rossi G.B.
        • Philipson L.
        • Sorrentino V.
        Regulation and expression of growth arrest-specific gene (gas-5) during growth, differentiation, and development.
        Mol. Cell. Biol. 1992; 12: 3514-3521
        • Schneider C.
        • King R.M.
        • Philipson L.
        Genes specifically expressed at growth arrest of mammalian cells.
        Cell. 1988; 54: 787-793
        • Smith C.M.
        • Steitz J.A.
        Classification of gas-5 as a multi-nucleolar-RNA (snoRNA) host gene and a member of the 5′-terminal oligopyrimidine gene family reveals common features of snoRNA host genes.
        Mol. Cell. Biol. 1998; 18: 6897-6909
        • Varnum B.C.
        • Young C.
        • Elliott G.
        • Garcia A.
        • Bartley T.D.
        • Fridell Y.W.
        • Hunt R.W.
        • Trall G.
        • Clogston C.
        • Toso R.J.
        • Yanagihara Y.
        • Bennett L.
        • Sylber M.
        • Merewether L.A.
        • Tseng A.
        • Escobar E.
        • Liu E.T.
        • Yamane H.K.
        Axl receptor tyrosine kinase stimulated by the vitamin K-dependent protein encoded by growth-arrest-specific gene 6.
        Nature. 1995; 373: 623-626
        • Manfioletti G.
        • Brancolini C.
        • Avanzi G.
        • Schneide C.
        The protein encoded by a growth arrest-specific gene (gas6) is a new member of the vitamin K-dependent proteins related to protein S, a negative coregulator in the blood coagulation cascade.
        Mol. Cell. Biol. 1993; 13: 4976-4985
        • Nakano T.
        • Higashino K.
        • Kikuchi N.
        • Kishino J.
        • Nomura K.
        • Fujita H.
        • Ohara O.
        • Arita H.
        Vascular smooth muscle cell-derived, Gla-containing growth-potentiating factor for Bs2+-mobilizing growth factors.
        J. Biol. Chem. 1995; 270: 5702-5705
        • Nakano T.
        • Kawamoto K.
        • Higashino K.
        • Arita H.
        Prevention of growth arrest-induced cell death of vascular smooth muscle cells by a product of growth arrest-specific gene, gas6.
        FEBS Lett. 1996; 387: 78-80
        • Bellosta P.
        • Costa M.
        • Lin D.A.
        • Basilico C.
        The receptor tyrosine kinase ARK mediates cell aggregation by nomophilic binding.
        Mol. Cell. Biol. 1995; 15: 614-625
        • Asnicar M.A.
        • Goheen M.
        • Bartlett M.B.
        • Smith J.W.
        • Lee C.-H.
        Upregulation of host mitochondrial ATPase 6 gene in Pneumocystic carinii-infected rat lungs.
        J. Euk. Microbiol. 1996; 43: 38S
        • Lamminen T.
        • Majander A.
        • Juvonen V.
        • Wikstrom M.
        • Aula P.
        • Nikoskelainen E.
        • Savontaus M.-L.
        A mitochondrial mutation at nt 9101 in the ATP synthase 6 gene associated with deficient oxidative phosphorylation in a family with Leber Hereditary Optic Neuroretinopathy.
        Am. J. Hum. Genet. 1995; 56: 1238-1240
        • Holt K.J.
        • Harding A.E.
        • Petty R.K.
        • Morgan-Hughes J.A.
        A new mitochondrial disease associated with mitochondrial DNA heteroplasmy.
        Am. J. Hum. Genet. 1990; 46: 428-433
        • Tatuch Y.
        • Christodoulou J.
        • Feigenbaum A.
        • Clarke J.T.R.
        • Wherret J.
        • Smith C.
        • Rudd N.
        • Petrova-Benedict R.
        • Robinson B.H.
        Heteroplasmic mtDNA mutation (T-G) at 8993 can cause Leigh Disease when the percentage of abnormal mtDNA is high.
        Am. J. Hum. Genet. 1992; 50: 852-858
        • Shoffner J.M.
        • Fernhoff P.M.
        • Krawiecki N.S.
        • Caplan D.B.
        • Holt P.J.
        • Koontz D.A.
        • Takei Y.
        • Newan N.J.
        • Oriz R.G.
        • Polak M.
        • Ballinger S.W.
        • Lott M.T.
        • Wallace D.C.
        Subacute necrotizing encephalopathy: oxidative phosphorylation defects and the ATPase 6 point mutation.
        Neurology. 1992; 42: 2168-2174
        • Trounce I.
        • Neill S.
        • Wallace D.C.
        Cytoplasmic transfer of the mtDNA at 9883 TG (ATP6) point mutation associated with Leigh syndrome into mtDNA-less cells demonstrates cosegregation with a decrease in state III respiration and ADP/O ratio.
        Proc. Natl. Acad. Sci. USA. 1994; 91: 8334-8338
        • Yang J.
        • Liu X.
        • Bhalla K.
        • Kim C.N.
        • Ibrado A.M.
        • Cai J.
        • Peng T.-I.
        • Jones D.P.
        • Wang X.
        Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked.
        Science. 1997; 275: 1129-1132
        • Higuchi M.
        • Aggarwal B.B.
        • Yeh E.T.H.
        Activation of CPP32-like protease in tumor necrosis factor-induced apoptosis is dependent on mitochondrial function.
        J. Clin. Invest. 1997; 99: 1751-1758
        • Bjorkerud S.
        • Bjorkerud B.
        Apoptosis is abundant in human atherosclerotic lesions, especially in inflammatory cells (macrophages and T cells), and may contribute to the accumulation of gruel and plaque instability.
        Am. J. Pathol. 1996; 149: 367-380