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• W2023841557 abstract "Vascular smooth muscle cells (SMC) synthesize insulin-like growth factor-I (IGF-I), which is a mitogen for this cell type in vitro. Since IGF binding proteins (IGFBP) modulate IGF bioactivity, we determined which IGFBPs were secreted by porcine SMC. Porcine SMC secreted 34,000 and 24,000 M(r) forms of IGFBPs which were identified as IGFBP-2 and IGFBP-4, respectively, by immunoblotting. Northern blot analysis showed single transcripts of 1.6 kb and 2.4 kb for IGFBP-2 and IGFBP-4, respectively. Secretion of IGFBP-2 was not regulated to a significant degree, with insulin, IGF-II, IGF-I, forskolin, and dibutyryl cyclic adenosine monophosphate (cAMP) inducing minimal changes in IGFBP-2 secretion of less than 30% by radioimmunoassay (RIA). Insulin increased (2.8 +/- 0.1-fold) the abundance of IGFBP-4 protein in conditioned media (CM) and increased IGFBP-4 mRNA levels. Growth factors for SMC such as platelet-derived growth factor (PDGF), fibroblast growth factor (FGF), epidermal growth factor (EGF), and transforming growth factor beta-1 (TGF beta-1) were without effect on either IGFBP-2 or -4. IGF-I treatment decreased the amount of IGFBP-4 present in CM, but a corresponding decrease in IGFBP-4 mRNA levels was not observed. In order to determine if IGFBP-4 could modulate IGF-I bioactivity, IGFBP-4 was added to pSMCs with and without IGF-I. IGF-I alone (20 ng/ml) induced a 1.6 to threefold increase in 3H-thymidine incorporation. Addition of IGFBP-4 (between 50 and 250 ng/ml) to cultures containing IGF-I (20 ng/ml) had no effect on DNA synthesis compared to that observed with IGF-I alone, while 500 ng/ml consistently caused a small decrease (15 +/- 5%; mean +/- SE). Immunoblotting of the CM obtained at the end of the 3H-thymidine assay showed a loss of intact IGFBP-4 in the cultures containing IGF-I. This corresponded with an increase in the abundance of a 16,000 M(r) immunoreactive fragment that did not bind IGF-I. Coincubation with insulin had no effect on the amount of IGFBP-4 that was converted to fragment, suggesting that the reaction was dependent upon IGF-I binding to IGFBP-4. In contrast, addition of IGFBP-4 (500 ng/ml) to human fibroblast cultures with IGF-I (20 ng/ml) almost completely inhibited the stimulatory effect of IGF-I on DNA synthesis and no increase in fragment was detected in the CM. In summary, SMC secrete IGFBP-2 and IGFBP-4, both of which have been shown to regulate IGF-mediated DNA synthesis.(ABSTRACT TRUNCATED AT 400 WORDS)" @default.
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• W2023841557 date "1993-10-01" @default.
• W2023841557 modified "2023-10-15" @default.
• W2023841557 title "Vascular smooth muscle cells synthesize two forms of insulin-like growth factor binding proteins which are regulated differently by the insulin-like growth factors" @default.
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• W2023841557 doi "https://doi.org/10.1002/jcp.1041570107" @default.
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