Effect of antisense mitogen-activated protein kinase oligonucleotides on rat vascular smooth muscle cell proliferation induced by EGF in vitro.
OBJECTIVE
To study the preventive effect of down-regulating mitogen-activated protein kinase (MAPK) on vascular smooth muscle cell (VSMC) proliferation.


METHODS
Cultured rat VSMC was pretreated with a phosphorothioate-protected 17-mer antisense MAPK oligodeoxynucleotides (ODN) directed against the initiation of translation sites of the p42- and p44-MAPK isoforms by liposomal transfection. A 17-mer sense and a random sequence MAPK ODN were used as control. After liposomal transfection, cells were exposed to epidermal growth factor (EGF) 1 nmol.L-1 for 10 min and then harvested in lysis buffer. MAPK activity was measured by Western blot and P-81 phosphocellulose filter papers method by using [gamma-32P] ATP and myelin basic protein as substrate. DNA synthesis was measured by [3H]thymidine incorporation.


RESULTS
Antisense ODN 0.2 mumol.L-1 reduced EGF-induced MAPK activities by 84%, and inhibited VSMC [3H]thymidine incorporation stimulated by EGF.


CONCLUSIONS
A 17-mer MAPK antisense oligonucleotide directed against the initiation of translation sites of the p44- and p42-MAPK inhibited EGF-stimulated rat VSMC proliferation.
Huang
S L
SL
Laboratory of Molecular Pharmacology, Hu-nan Medical University, Changsha, China.
Ding
B
B
Yu
Q S
QS
Guo
Z G
ZG
D000818:Animals; D001011:Aorta / Q000166:cytology; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000494:pharmacology*; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004815:Epidermal Growth Factor / Q000037:antagonists & inhibitors; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D020319:Oligodeoxyribonucleotides, Antisense; D004815:Epidermal Growth Factor; D017871:Calcium-Calmodulin-Dependent Protein Kinases