Soluble CD40 ligand induces human coronary artery smooth muscle cells proliferation and migration.
BACKGROUND
Clinical data have shown that an increased level of serum soluble CD40 ligand (sCD40L) is associated with atherosclerogenesis. We hypothesize that sCD40L induces proliferation and migration of vascular smooth muscle cells (VSMCs) through activation of matrix metalloproteinases (MMPs).


METHODS
Human VSMCs were treated with sCD40L (1 or 5 microg/mL). Cell proliferation and migration were studied using a nonradioactive cell proliferation assay (MTT) and a modified Boyden chamber combined with a scrape-wound assay, respectively. Messenger RNA (mRNA) and protein levels of MMP-2 and MMP-9 were measured with real-time polymerase chain reaction and enzyme-linked immunosorbent assays. Neutralizing antibodies against MMP-2 or MMP-9 were used to evaluate their effects on sCD40L-induced cell proliferation and migration.


RESULTS
MTT assay showed a 35% increase in cell proliferation in the high-dose (5 microg/mL) sCD40L-treated group. Cell migration was also increased by 33% (Transwell assay) to 3-fold (scrape-wound assay) after high-dose sCD40L treatment. When cells were treated with 5 microg/mL of sCD40L for 24 hours, significant decreases in MMP-2 and increases in MMP-9 mRNA and protein levels were observed. Neutralizing antibodies against MMP-9 effectively blocked sCD40L-induced cell proliferation and migration.


CONCLUSIONS
This study suggests that sCD40L increases VSMC proliferation and migration through the MMP-9 pathway, which may be a potential mechanism through which sCD40L induces intimal hyperplasia and atherosclerosis.
Chai
Hong
H
Division of Vascular and Endovascular Surgery, Department of Surgery, Stanford University, Stanford, Palo Alto, CA 94305, USA.
Aghaie
Kamran
K
Zhou
Wei
W
D050197:Atherosclerosis / Q000378:metabolism; D023201:CD40 Ligand / Q000494:pharmacology*; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000166:cytology* / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D006801:Humans; D006965:Hyperplasia / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D017539:Tunica Intima / Q000473:pathology
D012333:RNA, Messenger; D023201:CD40 Ligand; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9