PMID- 9747449 OWN - NLM STAT- MEDLINE DCOM- 19981015 LR - 20190512 IS - 0008-6363 (Print) IS - 0008-6363 (Linking) VI - 38 IP - 3 DP - 1998 Jun TI - Molecular cloning of genes differentially regulated by TNF-alpha in bovine aortic endothelial cells, fibroblasts and smooth muscle cells. PG - 802-13 AB - OBJECTIVE: Tumor necrosis factor-alpha (TNF-alpha) is a pleiotropic-cytokine binding to and thereby stimulating vascular cells. TNF-alpha mediated intermediate stimulation of vascular cells is believed to play a pivotal role in the development of arteriosclerosis. While extensive information has recently become available on gene induction by TNF-alpha, less is known about gene suppression by TNF-alpha in vascular cells. Endothelial cells are the first cell layer within the vessel wall interacting with circulating, cytokine releasing cells. Therefore, they were selected as target for these study. METHODS: A differential screening approach has been used to isolate cDNAs whose abundance was suppressed by incubating bovine aortic endothelial cells (BAEC) for 6 h with 1 nM TNF-alpha. The gene expression of 6 isolated cDNAs after TNF-alpha was investigated by dot blots and nuclear run-on analysis in BAEC. The investigated genes were partially or completely sequenced. Differential expression after TNF-alpha stimulation of BAEC, bovine fibroblasts and vascular smooth muscle cells (SMC) was studied by Northern blots. RNA transcripts of the clone C7 in aortic aneurysms were examined by in situ hybridization. RESULTS: 49 independent cDNAs were isolated by the differential screening approach and 6 clones were further analyzed. These genes were downregulated in a time and dose dependent manner in BAEC. Sequence analysis revealed that 3 cDNAs encoded previously unidentified genes (C1, C5, C7), while 3 encoded known genes: connective tissue growth factor (CTGF; A1), fibronectin (A8) and the mitochondrial genome (B1). A1 and B1 were suppressed in BAEC, fibroblasts and SMC, whereas A8, C1, C5 and C7 were not uniformly downregulated in the investigated cells. C7 RNA transcripts were exclusively induced in the endothelium of an uninflamed aortic aneurysm. The transcripts were undetectable in an inflamed aortic aneurysm and control vessels. CONCLUSIONS: Gene suppression is a prominent feature of the intermediate effect of TNF-alpha on endothelial cells. Differences in the expression of the tested genes in endothelial cells, fibroblasts and vascular smooth muscle cells open possibilities for the study of cellular interactions in the vascular wall in disease situations with high local TNF-alpha concentrations. FAU - Lin, J AU - Lin J AD - Department of Medicine I, University of Heidelberg, Germany. FAU - Liliensiek, B AU - Liliensiek B FAU - Kanitz, M AU - Kanitz M FAU - Schimanski, U AU - Schimanski U FAU - Bohrer, H AU - Bohrer H FAU - Waldherr, R AU - Waldherr R FAU - Martin, E AU - Martin E FAU - Kauffmann, G AU - Kauffmann G FAU - Ziegler, R AU - Ziegler R FAU - Nawroth, P P AU - Nawroth PP LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - England TA - Cardiovasc Res JT - Cardiovascular research JID - 0077427 RN - 0 (DNA, Complementary) RN - 0 (Tumor Necrosis Factor-alpha) SB - IM MH - Animals MH - Aorta MH - Blotting, Northern MH - Cattle MH - Cells, Cultured MH - Cloning, Molecular MH - DNA, Complementary/*genetics MH - Dose-Response Relationship, Drug MH - Endothelium, Vascular/drug effects/*metabolism MH - Fibroblasts/drug effects/metabolism MH - Gene Expression Regulation/*drug effects MH - In Situ Hybridization MH - Muscle, Smooth, Vascular/drug effects/metabolism MH - Sequence Analysis, DNA MH - Transcriptional Activation MH - Tumor Necrosis Factor-alpha/*pharmacology EDAT- 1998/09/25 00:00 MHDA- 1998/09/25 00:01 CRDT- 1998/09/25 00:00 PHST- 1998/09/25 00:00 [pubmed] PHST- 1998/09/25 00:01 [medline] PHST- 1998/09/25 00:00 [entrez] AID - S0008-6363(98)00055-8 [pii] AID - 10.1016/s0008-6363(98)00055-8 [doi] PST - ppublish SO - Cardiovasc Res. 1998 Jun;38(3):802-13. doi: 10.1016/s0008-6363(98)00055-8.