Endogenously generated nitric oxide by nitric-oxide synthase gene transfer inhibits cellular proliferation.
We investigated whether endothelial nitrite oxide synthase (NOS) gene transfer inhibited cellular proliferation. Endothelial NOS and endothelin type A receptor genes were transferred into 293 cells, a human embryonic kidney cell line, by calcium-phosphate coprecipitation. The cytosolic free Ca(2+) levels ([Ca(2+)](i)) of transfected cells were estimated with fura-2 fluorescence. Thymidine incorporation was increased by endothelin-1 in type A receptor-transfected cells. The endothelial NOS gene transfer did not affect endothelin-1-induced increase in [Ca(2+)](i) of type A receptor-transfected cells, but markedly inhibited mitogen-activated protein kinase and c-fos promoter activities. The endothelial NOS gene transfer also inhibited thymidine incorporation into type A receptor-transfected cells in response to endothelin-1, which was abolished in the presence of the NOS inhibitor N(G)-monomethyl-L-arginine acetate. The endothelin-1-induced increase in cell number was significantly suppressed by endothelial NOS gene transfer as well as by the mitogen-activated protein kinase inhibitor PD98059. These results indicate that endothelial NOS gene transfer inhibits cellular proliferation via inhibition of the mitogen-activated protein kinase cascade.
Maeda
Y
Y
Department of Cardiology, Institute of Hematology, Jichi Medical School, Tochigi, Japan.
Ikeda
U
U
Oya
K i
Ki
Shimpo
M
M
Ueno
S
S
Okada
K
K
Saito
T
T
Mano
H
H
Ozawa
K
K
Shimada
K
K
D002118:Calcium / Q000378:metabolism; D002455:Cell Division / Q000502:physiology*; D002478:Cells, Cultured; D004622:Embryo, Mammalian; D019332:Endothelin-1 / Q000494:pharmacology; D004727:Endothelium / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005419:Flavonoids / Q000494:pharmacology; D016257:Fura-2; D006801:Humans; D066298:In Vitro Techniques; D007668:Kidney / Q000502:physiology; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000096:biosynthesis / Q000502:physiology*; D019001:Nitric Oxide Synthase / Q000235:genetics*; D010710:Phosphates / Q000737:chemistry; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism; D017466:Receptors, Endothelin / Q000235:genetics*; D013936:Thymidine / Q000493:pharmacokinetics; D014162:Transfection
D019332:Endothelin-1; D004791:Enzyme Inhibitors; D005419:Flavonoids; D010710:Phosphates; D016760:Proto-Oncogene Proteins c-fos; D017466:Receptors, Endothelin; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D020928:Mitogen-Activated Protein Kinases; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one; D002118:Calcium; D016257:Fura-2; D019331:NG-Nitroarginine Methyl Ester; D013936:Thymidine