Abstract
We have demonstrated that a synthetic DNA enzyme targeting early growth response factor-1 (Egr-1) can inhibit neointimal hyperplasia following vascular injury. However, the detailed mechanism of this inhibition is not known. Thus, the objective of the present study was to further investigate potential inhibitory mechanisms. Catalytic DNA (ED5) and scrambled control DNA enzyme (ED5SCR) were synthesized and transfected into primary cultures of rat vascular smooth muscle cells (VSMCs). VSMC proliferation and DNA synthesis were analyzed by the MTT method and BrdU staining, respectively. Egr-1, TGF-beta1, p53, p21, Bax, and cyclin D1 expression was detected by RT-PCR and Western blot. Apoptosis and cell cycle assays were performed by FACS. Green fluorescence could be seen localized in the cytoplasm of 70.6 +/- 1.52 and 72 +/- 2.73% VSMCs 24 h after transfection of FITC-labeled ED5 and ED5SCR, respectively. We found that transfection with ED5 significantly inhibited cultured VSMC proliferation in vitro after 24, 48, and 72 h of serum stimulation, and also effectively decreased the uptake of BrdU by VSMC. ED5 specifically reduced serum-induced Egr-1 expression in VSMCs, further down-regulated the expression of cyclin D1 and TGF-beta1,...Continue Reading
References
Oct 1, 1992·The Journal of Clinical Investigation·S NikolL Weir
Sep 1, 1991·The Journal of Clinical Investigation·M W MajeskyM A Reidy
May 9, 1988·The American Journal of Cardiology·K M Kent
Sep 1, 1988·Journal of the American College of Cardiology·M NobuyoshiH Ohishi
Jun 15, 1984·The American Journal of Cardiology·D R HolmesK M Detre
Mar 1, 1994·The Journal of Clinical Investigation·Y G WolfE Ruoslahti
Feb 1, 1996·Circulation Research·R SarkarR C Webb
Mar 8, 1996·Science·L M KhachigianT Collins
Apr 29, 1997·Proceedings of the National Academy of Sciences of the United States of America·S W Santoro, G F Joyce
Aug 8, 1997·The Journal of Biological Chemistry·P NairV M Rangnekar
May 29, 1999·Circulation Research·J D SmithV Lindner
Nov 5, 1999·Nature Medicine·F S SantiagoL M Khachigian
Mar 11, 2000·The Journal of Clinical Investigation·T A McCaffreyT Collins
Apr 3, 2001·Cardiovascular Research·J ChamberlainD C Crossman
Oct 13, 2001·Circulation Research·H C LoweL M Khachigian
Nov 2, 2001·Atherosclerosis·S GoetzeR E Law
Mar 13, 2002·Circulation·Masazumi KumeKeizo Sugimachi
Mar 15, 2002·Gene Therapy·H YasumotoH Iwao
Oct 18, 2002·Journal of Cellular Physiology·Gerald Thiel, Giuseppe Cibelli
Jul 16, 2003·Circulation·Zakar H MnjoyanKen Fujise
Oct 15, 2003·Gene·Mingui FuYuqing E Chen
Nov 1, 2003·American Journal of Physiology. Heart and Circulatory Physiology·Antonio CurcioCiro Indolfi
Jan 9, 2004·Gene Therapy·K OhtaniA Takeshita
Jun 1, 2005·Life Sciences·Yuko TakahashiMitsuhiro Yokoyama
Feb 4, 2006·Circulation Research·Levon M Khachigian
Feb 5, 2008·International Journal of Cardiology·Gui-Nan LiuWu Yan