Inhibition of Growth of Human Gingival Fibroblasts by Chimeric DNA-RNA Hammerhead Ribozyme Targeting Transforming Growth Factor-β1
- Autores: Junko Yusa, Dr. Noboru Fukuda, Hideo Mugishima
- Localización: Journal of periodontology, ISSN 0022-3492, Vol. 76, Nº. 8, 2005, págs. 1265-1274
- Idioma: inglés
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Resumen
Background: Transforming growth factor (TGF)-β1 is involved in the pathogenesis of both drug-induced gingival overgrowth and hereditary gingival fibromatosis. Ribozymes enzymatically cleave target mRNAs and are expected to be utilized as the basis of novel nucleic acid-based therapies. We designed a chimeric DNA-RNA ribozyme targeting TGF-β1 mRNA and examined its effect on growth of gingival fibroblasts in culture.
Methods: Chimeric DNA-RNA hammerhead ribozyme with sequence complementary to the loop structure of human TGF-β1 mRNA was used. We evaluated transfer of the chimeric ribozyme by hemagglutinating virus of Japan (HVJ)-envelope into cultured human gingival fibroblasts in vitro and rat gingival tissues in vivo. We then examined effects of the chimeric ribozyme to TGF-β1 on proliferation and DNA synthesis in human gingival fibroblasts. We also examined effects of the chimeric ribozyme to TGF-β1 on expression of TGF-β1, type IV collagens, and fibronectin mRNAs and expression of TGF-β1 protein in human gingival fibroblasts.
Results: Chimeric ribozyme was sufficiently distributed into human fibroblasts in vitro and rat gingivae in vivo. Chimeric ribozyme to TGF-β1 significantly inhibited expression of TGF-β1, type IV collagen, and fibronectin mRNAs and TGF-β1 protein in human gingival fibroblasts. Mismatch ribozyme had no effect on expression of these molecules. Chimeric ribozyme to TGF-β1 also significantly inhibited proliferation and DNA synthesis in gingival fibroblasts.
Conclusion: Chimeric DNA-RNA ribozyme targeting TGF-β1 may be a useful gene therapy agent for treatment of gingival hyperplasia. J Periodontol 2005;76:1265-1274.
