Differential display identification of 40 genes with altered expression in activated human smooth muscle cells. Local expression in atherosclerotic lesions of smags, smooth muscle activation-specific genes.
Detailed knowledge on the molecular and cellular mechanisms that control (re)-differentiation of vascular smooth muscle cells (SMCs) is critical to understanding the pathological processes underlying atherogenesis. We identified by differential display/reverse transcriptase-polymerase chain reaction 40 genes with altered expression in cultured SMCs upon stimulation with the conditioned medium of activated macrophages. This set of genes comprises 10 known genes and 30 novel genes, which we call "smags" (for smooth muscle activation-specific genes). To determine the in vivo significance of these (novel) genes in atherogenesis, we performed in situ hybridization experiments on vascular tissue. Specifically, FLICE (Fas-associated death domain-like interleukin-1beta-converting enzyme)-like inhibitory protein (FLIP) is expressed in neointimal SMCs as well as in lesion macrophages and endothelial cells, whereas the expression of the novel genes smag-63, smag-64, and smag-84 is restricted to neointimal SMCs. Characterization of full-length smag-64 cDNA revealed that it encodes a novel protein of 66 amino acids. smag-82 cDNA comprises the complete, unknown, 3'-untranslated region of fibroblast growth factor-5. Collectively, our results illustrate the complex changes of SMC gene expression that occur in response to stimulation with cytokines and growth factors secreted by activated macrophages. Moreover, we identified interesting candidate genes that may play a role in the differentiation of SMCs during atherogenesis.
de Vries
C J
CJ
Departments of Biochemistry and Vascular Medicine, Academic Medical Center, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands. c.j.devries@amc.uva.nl
van Achterberg
T A
TA
Horrevoets
A J
AJ
ten Cate
J W
JW
Pannekoek
H
H
D000328:Adult; D001011:Aorta / Q000166:cytology; D001158:Arteries / Q000166:cytology; D001161:Arteriosclerosis / Q000209:etiology / Q000235:genetics* / Q000378:metabolism; D053446:CASP8 and FADD-Like Apoptosis Regulating Protein; D002352:Carrier Proteins / Q000096:biosynthesis; D002450:Cell Communication; D002454:Cell Differentiation / Q000235:genetics; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology; D018076:DNA, Complementary / Q000302:isolation & purification; D051521:Fibroblast Growth Factor 5; D005346:Fibroblast Growth Factors / Q000096:biosynthesis; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D007083:Iliac Artery / Q000166:cytology; D047908:Intracellular Signaling Peptides and Proteins; D008077:Lipoproteins, LDL; D008262:Macrophage Activation; D008264:Macrophages / Q000166:cytology / Q000502:physiology; D009124:Muscle Proteins / Q000096:biosynthesis / Q000235:genetics*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D010084:Oxidation-Reduction; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014470:Umbilical Cord / Q000098:blood supply / Q000166:cytology
D053446:CASP8 and FADD-Like Apoptosis Regulating Protein; C506386:CFLAR protein, human; D002352:Carrier Proteins; D017077:Culture Media, Conditioned; D018076:DNA, Complementary; C496449:FGF5 protein, human; D047908:Intracellular Signaling Peptides and Proteins; D008077:Lipoproteins, LDL; D009124:Muscle Proteins; D012333:RNA, Messenger; C099252:oxidized low density lipoprotein; D051521:Fibroblast Growth Factor 5; D005346:Fibroblast Growth Factors