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Article de Anglais | WPRIM | ID: wpr-147614

RÉSUMÉ

Latent transforming growth factor (TGF)-beta-binding protein (LTBP) is required for the assembly, secretion, matrix association, and activation of latent TGF-beta complex. To elucidate the cell specific expression of the genes of LTBP-1 and their splice variants and the factors that regulate the gene expression, we cultured primary human glomerular endothelial cells (HGEC) under different conditions. Basal expression of LTBP-1 mRNA was suppressed in HGEC compared to WI-38 human embryonic lung fibroblasts. High glucose, H2O2, and TGF-beta1 upregulated and vascular endothelial growth factor (VEGF) further downregulated LTBP-1 mRNA in HGEC. RT-PCR with a primer set for LTBP-1S produced many clones but no clone was gained with a primer set for LTBP-1L. Of 12 clones selected randomly, Sca I mapping and DNA sequencing revealed that only one was LTBP-1S and all the others were LTBP-1S delta 53. TGF-beta1, but not high glucose, H2O2 or VEGF, tended to increase LTBP-1S delta 53 mRNA. In conclusion, HGEC express LTBP-1 mRNA which is suppressed at basal state but upregulated by high glucose, H2O2, and TGF-beta1 and downregulated by VEGF. Major splice variant of LTBP-1 in HGEC was LTBP-1S delta 53. Modification of LTBP-1S delta 53 gene in HGEC may abrogate fibrotic action of TGF-beta1 but this requires confirmation.


Sujet(s)
Humains , Épissage alternatif , Séquence d'acides aminés , Lignée cellulaire , Cellules cultivées , Clonage moléculaire , Étude comparative , Cellules endothéliales/effets des médicaments et des substances chimiques , Régulation de l'expression des gènes , Glucose/pharmacologie , Peroxyde d'hydrogène/pharmacologie , Protéines et peptides de signalisation intracellulaire/génétique , Glomérule rénal/cytologie , Isoformes de protéines/génétique , ARN messager/génétique , RT-PCR , Transcription génétique , Transfection , Facteur de croissance transformant bêta/pharmacologie , Facteur de croissance endothéliale vasculaire de type A/pharmacologie
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