Skip Navigation

Cardiovascular Research 2000 48(1):138-147; doi:10.1016/S0008-6363(00)00157-7
© 2000 by European Society of Cardiology
This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow E-letters: Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when E-letters are posted
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Disclaimer
Google Scholar
Right arrow Articles by Teng, J.
Right arrow Articles by Kanmatsuse, K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Teng, J.
Right arrow Articles by Kanmatsuse, K.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Copyright © 2000, European Society of Cardiology

DNA–RNA chimeric hammerhead ribozyme to transforming growth factor-β1 mRNA inhibits the exaggerated growth of vascular smooth muscle cells from spontaneously hypertensive rats

Jian Teng, Noboru Fukuda*, Wen-Yang Hu, Mari Nakayama, Hirobumi Kishioka and Katsuo Kanmatsuse

Second Department of Internal Medicine, Nihon University School of Medicine, Tokyo 173-8610, Japan

* Corresponding author. Tel.: +81-3-3972-8111; fax: +81-3-3972-1098 nhukuda{at}med.nihon-u.ac.jp

Objective: The purpose of this study was to develop DNA–RNA chimeric hammerhead ribozyme against transforming growth factor-β1 (TGF-β1) mRNA as a gene therapy agent for arterial proliferative diseases. Methods: A 38-base hammerhead ribozyme against rat TGF-β1 mRNA, to produce cleavage at the GUC sequence at nucleotide 825 according to the secondary structure of rat TGF-β1 mRNA was designed. To enhance its stability, we synthesized a DNA–RNA chimeric ribozyme with two phosphorothioate linkages at the 3'-terminal. We also synthesized a mismatch ribozyme with single base change in the catalytic loop region as a control. These ribozymes were delivered into rat vascular smooth muscle cells (VSMC) from spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats by lipofectin-mediated transfection, and their biological effects were investigated. Results: According to in vitro cleavage studies, the synthetic ribozyme can cleave the synthetic substrate RNA into two RNA fragments. Chimeric ribozyme significantly inhibited DNA synthesis in VSMC from SHR but not in cells from WKY rats. Mismatch ribozyme showed only a little effect on growth of VSMC from SHR. Chimeric ribozyme significantly inhibited proliferation of VSMC from SHR; in contrast, the proliferation of VSMC from WKY rats was significantly increased by this chimeric ribozyme. Mismatch ribozyme did not affect proliferation of VSMC from either rat strain. Chimeric hammerhead ribozyme to rat TGF-β1 dose-dependently inhibited TGF-β1 mRNA expression detected by reverse transcription and polymerase chain reaction analysis in VSMC from both rat strains. Chimeric hammerhead ribozyme to rat TGF-β1 also dose-dependently inhibited TGF-β1 protein production detected by Western blot analysis. Conclusions: The present results demonstrated that our designed DNA–RNA chimeric hammerhead ribozyme to TGF-β1 mRNA might be a useful gene therapy agent for hypertensive vascular diseases.

KEYWORDS Gene therapy; Growth factors; Hypertension; Cell culture/isolation


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.