Novel E2F decoy oligodeoxynucleotides inhibit in vitro vascular smooth muscle cell proliferation and in vivo neointimal hyperplasia.
The transcription factor, E2F, plays a critical role in the trans-activation of several genes involved in cell cycle regulation. Previous studies showed that the transfection of cis element double-stranded decoy oligodeoxynucleotides (ODNs) corresponding to E2F binding sites inhibited the proliferation of vascular smooth muscle cells (VSMCs) and neointimal hyperplasia in injured vessels. We have developed a novel E2F decoy ODN with a circular dumbbell structure (CD-E2F) and compared its effects with those of the conventional phosphorothioated E2F decoy (PS-E2F) ODN. CD-E2F ODN was more stable than PS-E2F ODN, largely preserving its structural integrity after incubation in the presence of nucleases and sera. Moreover, CD-E2F ODN inhibited high glucose- and serum-induced transcriptional expression of cell cycle regulatory genes more strongly than PS-E2F ODN. Transfection of CD-E2F ODN resulted in more effective inhibition of VSMC proliferation in vitro and neointimal formation in vivo, compared with PS-E2F ODN. An approximately 40-50% lower dose of CD-E2F ODN than PS-E2F ODN was sufficient to attain similar effects. In conclusion, our results indicate that CD-E2F ODN may be a valuable tool in gene therapy protocols for inhibiting VSMC proliferation and studying transcriptional regulation.
Ahn
J D
JD
Department of Microbiology, Kyungpook National University, Taegu, Korea.
Morishita
R
R
Kaneda
Y
Y
Kim
H S
HS
Chang
Y-C
YC
Lee
K-U
KU
Park
J-Y
JY
Lee
H W
HW
Kim
Y-H
YH
Lee
I-K
IK
D000818:Animals; D001665:Binding Sites; D016893:Carotid Stenosis / Q000473:pathology / Q000628:therapy*; D018797:Cell Cycle Proteins; D002455:Cell Division; D002478:Cells, Cultured; D019925:Cyclin A / Q000235:genetics; D004268:DNA-Binding Proteins; D050684:E2F Transcription Factors; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D006965:Hyperplasia; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000378:metabolism / Q000473:pathology*; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D014157:Transcription Factors / Q000235:genetics*; D014162:Transfection; D017539:Tunica Intima / Q000473:pathology
D018797:Cell Cycle Proteins; D019925:Cyclin A; D004268:DNA-Binding Proteins; D050684:E2F Transcription Factors; D018809:Proliferating Cell Nuclear Antigen; D014157:Transcription Factors