No influence of OPG and its ligands, RANKL and TRAIL, on proliferation and regulation of the calcification process in primary human vascular smooth muscle cells.
The aim of this study was to examine the effects of the OPG-RANKL-TRAIL system on proliferation, regulation of calcification-associated genes and calcification of human vascular smooth muscle cells (HVSMCs). Small interfering (si)RNA-mediated knockdown of OPG was followed by treatment of HVSMCs with recombinant RANKL or TRAIL. Regulation of a calcification-associated gene set was assayed by pathway analysis of microarray results. The lack of OPG in HVSMCs or treatment with RANKL or TRAIL did not affect proliferation of HVSMCs. In addition, OPG, RANKL or TRAIL did not modify the regulation of a calcification-associated gene set. Finally, in the long term calcification assay, we found that cells isolated from seven different human donors showed a great variability in the response to RANKL and insulin. However, overall RANKL and/or insulin did not affect the development of calcification of HVSMCs. These studies indicate that OPG knockdown does not alter the calcification process in HVSMCs.
Department of Cardiovascular and Renal Research, Institute of Molecular Medicine, University of Southern Denmark, Denmark. molesen@health.sdu.dk
Malene
M
Olesen
Vibe
V
Skov
Mie
M
Mechta
Birgitte Hviid
BH
Mumm
Lars Melholt
LM
Rasmussen
D049109:Cell Proliferation; D002478:Cells, Cultured; D055785:Gene Knockdown Techniques; D006801:Humans; D007328:Insulin / Q000502:physiology; D009131:Muscle, Smooth, Vascular / Q000473:pathology; D032389:Myocytes, Smooth Muscle / Q000378:metabolism / Q000473:pathology* / Q000502:physiology; D020411:Oligonucleotide Array Sequence Analysis; D053244:Osteoprotegerin / Q000235:genetics / Q000378:metabolism*; D053245:RANK Ligand / Q000502:physiology*; D034622:RNA Interference; D053221:TNF-Related Apoptosis-Inducing Ligand / Q000502:physiology*; D059467:Transcriptome; D061205:Vascular Calcification / Q000235:genetics* / Q000378:metabolism
D007328:Insulin; D053244:Osteoprotegerin; D053245:RANK Ligand; D053221:TNF-Related Apoptosis-Inducing Ligand; C506030:TNFRSF11B protein, human; C505653:TNFSF10 protein, human; C505688:TNFSF11 protein, human