Adenovirus-mediated transfer of the atrial natriuretic peptide gene in rat pulmonary vascular smooth muscle cells leads to apoptosis.
Atrial natriuretic peptide (ANP) exhibits relaxant and growth-inhibiting effects on vascular smooth muscle cells (VSMCs). To obtain ANP gene expression in VSMCs, we built a recombinant adenovirus containing the ANP cDNA controlled by the adenovirus major late promotor (AdMLP-ANP). After pulmonary VSMC treatment with AdMLP-ANP at a multiplicity of infection ranging from 5 to 100 TCID(50)/cell, immunoreactive ANP was detectable in the cell culture medium at a level that reached 101 +/- 27 pmol/well after 2 days. The newly expressed ANP was biologically active, as evidenced by its ability to induce cyclic guanosine monophosphate accumulation in target cells and to mimic the effect of exogenous ANP (10(-8) to 10(-7) mol/L). Cell growth and survival of AdMLP-ANP-infected cells were decreased and were associated with the promotion of VSMC apoptosis. These effects, which occurred at a multiplicity of infection of 10 to 100 TCID(50)/cell, were observed neither in cells infected with the control adenoviral constructs (AdMLP-betaGAL and AdMLP-gD) nor in cells treated with exogenous ANP (10(-7) to 10(-6) mol/L). These results showing VSMC apoptosis in response to ANP gene expression may have important implications for the prevention of vascular remodeling by gene therapy.
Deprez
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Département de Physiologie et INSERM U296, Faculté de médecine, CHU Henri-Mondor, Créteil, France.
Darmon
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ME
Hira
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Adam
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M
Sanquer
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S
Teiger
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E
Chetboul
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V
Eloit
M
M
Adnot
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Pham
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D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D009320:Atrial Natriuretic Factor / Q000235:genetics*; D002455:Cell Division / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D004274:DNA, Recombinant; D005822:Genetic Vectors; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000502:physiology; D011651:Pulmonary Artery / Q000166:cytology; D051381:Rats
D004274:DNA, Recombinant; D009320:Atrial Natriuretic Factor