Effects of antisense peptide nucleic acid to platelet-derived growth factor A-chain on growth of vascular smooth muscle cells.
To investigate antisense peptide nucleic acid (PNA) as a gene therapy for the arterial proliferative diseases, the authors designed and examined the effects of an antisense PNA targeting platelet-derived growth factor (PDGF) A-chain on expression of PDGF A-chain and growth of vascular smooth muscle cells (VSMCs) from spontaneously hypertensive rats. A 15-mer antisense PNA complementary to the initiation codon of rat and human PDGF A-chain mRNA was synthesized and purified by high-performance liquid chromatography. Gel-shift assay and biomolecular interaction analysis (BIAcore) revealed that the antisense PNA bound weakly to the target RNA, whereas it bound strongly to the target DNA. Fluorescein-isothiocyanate-labeled antisense PNA to PDGF A-chain was taken up slowly and maintained in VSMCs for a prolonged period of time. Antisense PNA inhibited expression of PDGF A-chain mRNA and protein as well as DNA synthesis in VSMCs in a dose-independent manner. Inhibition of DNA synthesis by the antisense PNA was greater than that by the antisense DNA at a low concentration (0.5 micromol/L). These results suggest that antisense PNA to PDGF A-chain will be used as a gene therapy for vascular proliferative diseases such as hypertensive vascular diseases, restenosis of coronary arteries after angioplasty, and atherosclerosis.
Fukuda
Noboru
N
Second Department of Internal Medicine, Nihon University School of Medicine, Ooyaguchi-kami 30-1, Itabashi-ku, Tokyo 173-8610, Japan. fukudan@med.nihon-u.ac.jp
Furuya
Rie
R
Kishioka
Hirobumi
H
Suzuki
Ryo
R
Matsuda
Hiroyuki
H
Tahira
Yoshiko
Y
Takagi
Hiroto
H
Ikeda
Yukihiro
Y
Saito
Satoshi
S
Matsumoto
Koichi
K
Kanmatsuse
Katsuo
K
D000818:Animals; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000254:growth & development; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis / Q000494:pharmacology*; D020135:Peptide Nucleic Acids / Q000138:chemical synthesis / Q000494:pharmacology*; D010982:Platelet-Derived Growth Factor / Q000187:drug effects*; D051381:Rats; D011918:Rats, Inbred SHR; D020133:Reverse Transcriptase Polymerase Chain Reaction
D016376:Oligonucleotides, Antisense; D020135:Peptide Nucleic Acids; D010982:Platelet-Derived Growth Factor; C075278:platelet-derived growth factor A