RESUMO
We established two cell lines of human smooth muscle cells (SMC) by transfection of cells from the aortic intima and aortic media with origin-minus simian virus 40 (ori-minus SV40) DNA. Ori-minus SV40 DNA very efficiently immortalized human smooth muscle cells in culture. Proteins that these cell lines produced included type I, III, IV, and V collagens, fibronectin, and human matrix metalloproteinases (MMP)-1 (tissue collagenase), -2 ("type IV collagenase"), and -3 (stromelysin). The protein production in these cell lines generally mimicked that of normal SMC, but the immortalization stimulated the cell line of medial SMC to produce excessive MMP-2 and to secrete MMP-9 (92-kDa gelatinase). However, since these cell lines did not show a fully malignant phenotype, we concluded that, in addition to the degradation of extracellular matrix macromolecules, including basement membrane components by MMP-2, -3, and/or -9, some additional factors must be involved for the malignancy of fully transformed cells and that these immortalized human aortic SMC, which share many characteristics with normal SMC, will prove useful to study the role(s) of metalloproteinases in atherosclerosis.
Assuntos
Linhagem Celular Transformada , DNA Viral/genética , Músculo Liso Vascular/citologia , Vírus 40 dos Símios/genética , Animais , Antígenos Transformantes de Poliomavirus/análise , Aorta , Colágeno/biossíntese , Colagenases/biossíntese , Fibronectinas/biossíntese , Humanos , Metaloproteinase 1 da Matriz , Metaloproteinase 3 da Matriz , Metaloendopeptidases/biossíntese , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Músculo Liso Vascular/metabolismo , Neoplasias Experimentais/etiologia , Fenótipo , Transfecção , Ensaio Tumoral de Célula-Tronco , Proteína Supressora de Tumor p53/análiseRESUMO
The effect of linoleic acid hydroperoxide on in vitro production of matrix metalloproteinases (MMPs) by human skin fibroblasts was studied. The addition of linoleic acid hydroperoxide significantly increased the production of MMP-1 (tissue collagenase) and MMP-3 (stromelysin), while it rather decreased that of MMP-2 (gelatinase of 72 kDa; so-called "type IV collagenase"). The effect of lipid peroxides to alter collagen metabolism was discussed from pathogenic points of view.
Assuntos
Ácidos Linoleicos/farmacologia , Peróxidos Lipídicos/farmacologia , Metaloendopeptidases/biossíntese , Pele/efeitos dos fármacos , Células Cultivadas , Eletroforese em Gel de Poliacrilamida , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Humanos , Immunoblotting , Microscopia de Fluorescência , Pele/citologia , Pele/metabolismoRESUMO
The effect of linoleic acid hydroperoxide on replication of adenovirus type 2 DNA in cultured endothelial cells of bovine aorta was studied to elucidate the deleterious effect of lipid peroxides on replication of DNA in the arterial endothelial cells. When, just after the infection, the cells were incubated with 1 nmol (measured by the thiobarbituric acid reaction and expressed in terms of malondialdehyde) of the hydroperoxide, the replication of adenovirus type 2 DNA was markedly reduced. The level of the messenger RNA for adenovirus type 2 DNA polymerase was also reduced. In accordance with the reduction of the messenger RNA, the synthesis of the polymerase in the cells was reduced. The inhibition of DNA replication in the arterial endothelial cells by lipid peroxides was discussed in relation to atherogenesis.