[Suppression of vascular smooth muscle cell proliferation by an antisense oligonucleotide against PDGF receptor].
Percutaneous transluminal coronary angioplasty (PTCA) is widely accepted as a treatment of ischemic heart diseases, such as Angina Pectoris and Acute Myocardial Infarction. However, coronary restenosis frequently observed in the patients treated with PTCA has been one of the major drawbacks of this procedure. Although the pathogenesis involved in this coronary restenosis is not fully understood, multiple growth factors promoting vascular smooth muscle cell (VSMC) proliferation and migration are likely to play a pivotal role in this process. The purpose of this study is to suppress the platelet-derived growth factor (PDGF)-induced VSMC growth by antisense oligonucleotides against PDGF receptor mRNA. Three different antisense S-oligonucleotides (AS-No. 1-3) were synthesized around the initiation codon of the coding sequence of rat PDGF alpha receptor. Rat cultured VSMCs (A10) were incubated with the AS for 3 days, then exposed to 10ng/ml PDGF-AB for 24 hours. Subsequently, DNA synthesis was assessed by 3H-thymidine incorporation into the cells. Among three AS we tested, AS-No. 3 most effectively suppressed the PDGF-induced DNA synthesis. The DNA synthesis of A10 cells induced by 3% fatal bovine serum (FBS), 10ng/ml PDGF-AB and PDGF-BB were significantly inhibited by 1 microM AS-No. 3 (51%, 44% and 49%, respectively). Serum and PDGF-induced DNA synthesis was inhibited dose dependently by AS-No. 3 (0.25-1 microM range). Northern blot analysis revealed high level expression of PDGF beta receptor mRNA, but failed to detected alpha receptor message in control A10 cells. A quantitative RT-PCR method was used so as to detect subtle change of alpha receptor message, and the method showed that the expression of alpha receptor message was significantly suppressed in the cells treated by 1 microM AS-No. 3. In this study, antisense oligonucleotide targeting rat PDGF alpha receptor effectively suppressed the expression of the receptor, and inhibited PDGF/serum - induced DNA synthesis in rat vascular smooth muscle cells. In addition, these results provide an important impetus to initiating in vivo studies to determine the feasibility of antisense strategies in the prevention of coronary restenosis.
Sugiki
H
H
Department of Cardiovascular Medicine, Hokkaido University School of Medicine, Sapporo, Japan.
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D051381:Rats; D017479:Receptors, Platelet-Derived Growth Factor / Q000037:antagonists & inhibitors*
D016376:Oligonucleotides, Antisense; D004247:DNA; D017479:Receptors, Platelet-Derived Growth Factor