Hammerhead ribozyme targeting human platelet-derived growth factor A-chainmRNA inhibited the proliferation of human vascular smooth muscle cells

Platelet-derived growth factor (PDGF) A-chain contributes to the pathogenes is of cardiovascular proliferative diseases, such as hypertensive vascular disease, atherosclerosis, and re-stenosis of an artery after angioplasty. T o develop a ribozyme against human PDGF A-chain mRNA as a gene therapy for human arterial proliferative diseases, we designed and synthesized a 38-bas e hammerhead ribozyme to cleave human PDGF A-chain mRNA at the GUC sequence at nucleotide 591. In the presence of MgCl2, synthetic hammerhead ribozyme to human PDGF A-chain mRNA cleaved the synthetic target RNA to two RNA fra gments at a predicted size. Doses of 0.01-1.0 muM hammerhead ribozyme to hu man PDGF A-chain mRNA significantly inhibited angiotensin II (Ang If) and t ransforming growth factor (TGF)-beta (1)-induced DNA synthesis in vascular smooth muscle cells (VSMC) from human in a dose-dependent manner. One micro molor of hammerhead ribozyme to human PDGF A-chain mRNA significantly inhib ited Ang II-induced PDGF A-chain mRNA and PDGF-AA protein expressions in VS MC from humans. These results indicate that the designed hammerhead ribozym e to human PDGF A-chain mRNA effectively inhibited growth of human VSMC by cleaving the PDGF A-chain mRNA and inhibiting the PDGF-AA protein expressio n in human VSMC. This suggests that the designed hammerhead ribozyme to PDG F A-chain mRNA is a feasible gene therapy for treating arterial proliferati ve diseases. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.