Adenovirus-mediated gene transfer of C-type natriuretic peptide causes G1 growth inhibition of cultured vascular smooth muscle cells.
We have proposed the "vascular natriuretic peptide system", in which C-type natriuretic peptide (CNP), the third member of the natriuretic peptide family, can control vascular tone and growth as an endothelium-derived relaxing peptide. We aimed at overexpression of CNP gene in vascular smooth muscle cells (SMCs) by adenovirus-mediated gene transfer to examine the growth characteristics of SMCs via the augmentation of cGMP production. Rat aortic SMCs infected with Ad.CNP, a replication-deficient adenovirus driving rat CNP cDNA, produced 162 +/- 55 fmol/mL of CNP, which was 4,000 times higher than that produced by endothelial cells. cGMP production was also augmented in Ad.CNP-infected SMCs (2200 +/- 270 fmol/10(4) cells). Accordingly, significant growth inhibition was observed in SMCs infected with Ad.CNP. The flow cytometry analysis revealed that the population of the S and G2 + M phases was reduced by 60% of the control in Ad.CNP-infected SMCs. The gene expression of ANP-B receptor, which is expressed abundantly in SMCs with the synthetic phenotype, was suppressed in Ad.CNP-infected SMCs, while the gene expression of ANP-A receptor, which is expressed predominantly in SMCs with the contractile phenotype, became detectable in Ad.CNP-infected SMCs. In addition, the gene expression of smooth muscle myosin heavy chain-2 (SM-2), which is the molecular marker of highly-differentiated SMCs, was also induced in Ad.CNP-treated SMCs. These results suggest that cGMP cascade activation induces re-differentiation of SMCs. The present study demonstrated that overexpression of CNP induced growth inhibition of SMCs at the G1 phase with possible alteration of the phenotype.
Doi
K
K
Department of Medicine and Clinical Science, Kyoto University Graduate School of Medicine, Japan.
Itoh
H
H
Ikeda
T
T
Hosoda
K
K
Ogawa
Y
Y
Igaki
T
T
Yamashita
J
J
Chun
T H
TH
Inoue
M
M
Masatsugu
K
K
Matsuda
K
K
Ohmori
K
K
Nakao
K
K
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001013:Aorta, Thoracic; D009320:Atrial Natriuretic Factor / Q000096:biosynthesis / Q000235:genetics*; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000378:metabolism; D019925:Cyclin A / Q000096:biosynthesis / Q000235:genetics; D016193:G1 Phase / Q000235:genetics*; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D006131:Growth Inhibitors / Q000235:genetics*; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D020098:Natriuretic Peptide, C-Type; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats
D019925:Cyclin A; D006131:Growth Inhibitors; D011506:Proteins; D012333:RNA, Messenger; D020098:Natriuretic Peptide, C-Type; D009320:Atrial Natriuretic Factor; D006152:Cyclic GMP