TY - JOUR T1 - Synthetic Pyrrole-Imidazole Polyamide Inhibits Expression of the Human Transforming Growth Factor-β1 Gene JF - Journal of Pharmacology and Experimental Therapeutics JO - J Pharmacol Exp Ther SP - 571 LP - 575 DO - 10.1124/jpet.105.089086 VL - 315 IS - 2 AU - Yu-Mu Lai AU - Noboru Fukuda AU - Takahiro Ueno AU - Hiroyuki Matsuda AU - Satoshi Saito AU - Koichi Matsumoto AU - Hirohito Ayame AU - Toshikazu Bando AU - Hiroshi Sugiyama AU - Hideo Mugishima AU - Kazuo Serie Y1 - 2005/11/01 UR - http://jpet.aspetjournals.org/content/315/2/571.abstract N2 - Pyrrole-imidazole (Py-Im) polyamides can bind to the predetermined base pairs in the minor groove of double-helical DNA with high affinity. These synthetic small molecules can interfere with transcription factor-DNA interaction and inhibit or activate the transcription of corresponding genes. In the present study, we designed and synthesized a Py-Im polyamide to target -545 to -539 base pairs of human transforming growth factor-β1 (hTGF-β1) promoter adjacent to the fat-specific element 2 (FSE2) to inhibit the expression of the gene. Gel mobility shift assay showed that the synthetic Py-Im polyamide binds to its corresponding double-strand oligonucleotides, whereas the mismatch polyamides did not bind. Fluorescein isothiocyanate-labeled Py-Im polyamide was detected in the nuclei of human vascular smooth muscle cells (VSMCs) after 2- to 48-h incubation. Py-Im polyamide significantly decreased the promoter activity of hTGF-β1 determined by in vitro transcription experiments and luciferase assay. In cultured human VSMCs, Py-Im polyamide targeting hTGF-β1 promoter significantly inhibited expressions of hTGF-β1 mRNA and protein. These results indicate that the synthetic Py-Im polyamide designed to bind hTGF-β1 promoter inhibited hTGF-β1 gene and protein expression successfully. This novel agent will be used for the TGF-β-related diseases as a gene therapy. The American Society for Pharmacology and Experimental Therapeutics ER -