Ribozyme to human TGF-beta1 mRNA inhibits the proliferation of human vascular smooth muscle cells.
Transforming growth factor-beta (TGF-beta) has been reported to be involved in the pathogenesis of cardiovascular proliferative diseases such as hypertensive vascular disease, atherosclerosis, and arterial restenosis after angioplasty. We designed a 38-base DNA-RNA chimeric hammerhead ribozyme to cleave human TGF-beta1 mRNA as a gene therapy for human arterial proliferative diseases. In the presence of MgCl(2), synthetic ribozyme to human TGF-beta1 mRNA cleaved the synthetic target RNA into two RNA fragments of predicted size. A control mismatch ribozyme, with one different base in the catalytic loop region, was inactive. DNA-RNA chimeric ribozyme (0. 01-1.0 microM) significantly inhibited angiotensin II (Ang II)-stimulated DNA synthesis in a dose-dependent manner in human vascular smooth muscle cells (VSMC). The mismatch ribozyme did not affect Ang II-stimulated DNA synthesis in the cells. DNA-RNA chimeric ribozyme (1.0 microM) inhibited the proliferation of human VSMC in the presence of Ang II. DNA-RNA chimeric ribozyme (1.0 microM) significantly inhibited Ang II-stimulated TGF-beta1 mRNA and protein expression in human VSMC. These results indicate that the designed DNA-RNA chimeric hammerhead ribozyme targeted to human TGF-beta1 mRNA can effectively and potentially inhibit growth of human VSMC by cleaving the TGF-beta1 mRNA. This finding suggests that this ribozyme will be useful in the gene therapy of arterial proliferative diseases.
Su
J Z
JZ
Second Department of Internal Medicine, Nihon University School of Medicine, Tokyo, 173-8610, Japan.
Fukuda
N
N
Hu
W Y
WY
Kanmatsuse
K
K
D001483:Base Sequence; D002455:Cell Division / Q000502:physiology*; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D009690:Nucleic Acid Conformation; D016337:RNA, Catalytic / Q000502:physiology*; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics*; D016212:Transforming Growth Factor beta / Q000235:genetics*
D016337:RNA, Catalytic; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta