title: Endothelial monocyte activating polypeptide-II induced gene expression changes in endothelial cells.
abstract: In the current study we used microarray (MA) analysis to examine gene expression changes in human umbilical vein endothelial cells (HUVEC) exposed to the tumor-derived cytokine, endothelial monocyte-activating polypeptide-II (EMAP-II). HUVEC treated with EMAP-II for 0.5, 1, 2, 4 and 8 h, were analyzed using 10K cDNA arrays. Our results demonstrated that changes in gene expression of <0.5 and >2 fold were seen for 69 genes and the majority of gene changes occurred early. Validation of MA analysis for 10 genes by real time RT-PCR, demonstrated the gene changes to be consistent and specific to HUVEC when compared to human fibroblasts treated with EMAP-II. Among these genes, downregulated in ovarian cancer 1 (DOC1) gene was studied further because of its possible role in EMAP-II induced cytoskeletal remodeling. DOC1 expression was silenced using small interfering RNA. SiRNA to DOC1 completely abolished EMAP-II stimulated gene expression of DOC1. Silencing of DOC1 gene expression reversed the modulatory effect of EMAP-II on 4 other genes, suggesting that DOC1 might play a role in mediating some of the effects of EMAP-II on endothelial cells.
authors.affiliation: Surgery Branch, Center for Cancer Research, National Cancer Institute, Bethesda, MD 20892, USA.
authors.forename: Anita T
authors.initials: AT
authors.lastname: Tandle
authors.forename: Chiara
authors.initials: C
authors.lastname: Mazzanti
authors.forename: H Richard
authors.initials: HR
authors.lastname: Alexander
authors.forename: David D
authors.initials: DD
authors.lastname: Roberts
authors.forename: Steven K
authors.initials: SK
authors.lastname: Libutti
mesh_terms: D049109:Cell Proliferation; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D018076:DNA, Complementary / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism; D004730:Endothelium, Vascular / Q000166:cytology; D005347:Fibroblasts / Q000378:metabolism; D005786:Gene Expression Regulation; D020868:Gene Silencing; D006801:Humans; D007091:Image Processing, Computer-Assisted; D047908:Intracellular Signaling Peptides and Proteins; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000502:physiology*; D020411:Oligonucleotide Array Sequence Analysis; D034741:RNA, Small Interfering / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000096:biosynthesis / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012542:Scattering, Radiation; D013997:Time Factors; D014471:Umbilical Veins / Q000166:cytology
chemical_list: D016207:Cytokines; D018076:DNA, Complementary; C504459:FILIP1L protein, human; D047908:Intracellular Signaling Peptides and Proteins; D009363:Neoplasm Proteins; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; C077219:small inducible cytokine subfamily E, member 1