title: Green fluorescent protein selectively induces HSP70-mediated up-regulation of COX-2 expression in endothelial cells.
abstract: Reporter genes, including green fluorescent protein (GFP), have been used to monitor the expression of transgenes introduced into vascular cells by gene transfer vectors. Here, we demonstrate that GFP by itself can selectively induce expression of certain genes in endothelial cells. Elevation of the cytoplasmic concentration of GFP in endothelial cells, specifically, resulted in a robust upregulation of heat shock protein 70 (HSP70). GFP induced both mRNA and protein expression of HSP70 in a dose-dependent manner. GFP-mediated up-regulation of HSP70 resulted in induction of cyclooxygenase-2 (COX-2) followed by prostaglandin E2 (PGE2) production. GFP-mediated up-regulation of HSP70 is independent of mitogen-activated protein kinase and phosphatidylinositol-3-kinase signaling cascades because inhibition of these pathways had no effect on HSP70 increases. Adenoviral delivery of GFP into murine vasculature significantly enhanced blood flow, suggesting that sufficient PGE2 is produced to induce vasodilation. Identification of the molecular partners that interact with GFP will increase our understanding of the vascular-specific factors that regulate stress angiogenesis and hemostasis.
authors.affiliation: Division of Hematology-Oncology and Genetic Medicine, Weill Medical College of Cornell University, 1300 York Ave, Rm D601, New York, NY 10021, USA.
authors.forename: Fan
authors.initials: F
authors.lastname: Zhang
authors.forename: Neil R
authors.initials: NR
authors.lastname: Hackett
authors.forename: George
authors.initials: G
authors.lastname: Lam
authors.forename: Joseph
authors.initials: J
authors.lastname: Cheng
authors.forename: Robert
authors.initials: R
authors.lastname: Pergolizzi
authors.forename: Lan
authors.initials: L
authors.lastname: Luo
authors.forename: Sergey V
authors.initials: SV
authors.lastname: Shmelkov
authors.forename: Jay
authors.initials: J
authors.lastname: Edelberg
authors.forename: Ronald G
authors.initials: RG
authors.lastname: Crystal
authors.forename: Shahin
authors.initials: S
authors.lastname: Rafii
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002478:Cells, Cultured; D051546:Cyclooxygenase 2; D015232:Dinoprostone / Q000096:biosynthesis; D004431:Ear, External / Q000098:blood supply; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D007202:Indicators and Reagents / Q000378:metabolism*; D007527:Isoenzymes / Q000235:genetics*; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism*; D020935:MAP Kinase Signaling System / Q000502:physiology; D008565:Membrane Proteins; D051379:Mice; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics*; D014158:Transcription, Genetic / Q000502:physiology; D014471:Umbilical Veins / Q000166:cytology; D015854:Up-Regulation / Q000502:physiology; D014664:Vasodilation / Q000502:physiology
chemical_list: D018840:HSP70 Heat-Shock Proteins; D007202:Indicators and Reagents; D007527:Isoenzymes; D008164:Luminescent Proteins; D008565:Membrane Proteins; D049452:Green Fluorescent Proteins; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011451:Prostaglandin-Endoperoxide Synthases; D019869:Phosphatidylinositol 3-Kinases; D015232:Dinoprostone