TGF-β stimulates collagen (I) in vascular smooth muscle cells via a short element in the proximal collagen promoter
TGF-β stimulates collagen (I) in vascular smooth muscle cells via a short element in the proximal collagen promoter
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DOI：
10.1016/s0022-4804(02)00037-9
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发表时间：
2003-01-01
影响因子：
2.2
通讯作者：
Liu, B
中科院分区：
文献类型：
--
作者：
Kubota, K;Okazaki, J;Liu, B
背景。细胞外基质的积累有助于内膜增生的发展。转化生长因子β (tgf - β)刺激血管平滑肌细胞（SMC）中几种基质蛋白的产生，包括I型胶原，但tgf - β作用的潜在机制尚不清楚。材料和方法。tgf - β对I型胶原生物合成的影响通过[H-3]脯氨酸掺入试验和Northern blotting测定。采用瞬时转染法和凝胶迁移位移法对人α 2(I)前胶原蛋白（COL1A2）基因启动子进行了分析。用tgf - β治疗人血管SMC刺激胶原合成并增加a2(1)胶原mRNA水平。将人COL1A2启动子与萤火虫荧光素酶基因连接构建胶原-荧光素酶报告基因，在人SMC中瞬间表达。tgf - β治疗显著刺激了这种胶原-荧光素酶报告蛋白的活性。通过缺失分析，我们在人COL1A2启动子中发现了一个150 bp的DNA片段（- 334至- 184），作为tgf - β介导人SMC中胶原基因表达的位点。凝胶迁移迁移实验表明，这一150 bp的DNA片段与来自SMC的多个核因子形成了共轭物，tgf - β进一步增强了这一过程。tgf - β通过位于其启动子近端区域的DNA元件刺激人类I型胶原基因。干扰这种DNA元素和核因子之间相互作用的干预措施可能会阻止胶原蛋白的产生，从而可能对内膜增生的发展产生重大影响。(C) 2003 Elsevier Science（美国）
Background. Accumulation of extracellular matrix contributes to the development of intimal hyperplasia. Transforming growth factor beta (TGF-beta) stimulates the production of several matrix proteins in vascular smooth muscle cells (SMC) including type I collagen, but the underlying mechanisms of TGF-beta's effects are not well understood.Material and methods. The effect of TGF-beta on type I collagen biosynthesis was determined by a [H-3]proline incorporation assay and Northern blotting. The promoter of human alpha2(I) procollagen (COL1A2) gene was analyzed by transient transfection analysis and gel mobility shift assay.Results. Treatment of human vascular SMC with TGF-beta stimulated collagen synthesis and increased the level of a2(1) collagen mRNA. A collagen-luciferase reporter gene, constructed by linking the human COL1A2 promoter with the firefly luciferase gene, was transiently expressed in human SMC. Treatment with TGF-beta significantly stimulated the activity of this collagen-luciferase reporter. Using deletion analysis, we identified a 150 bp DNA fragment (- 334 to - 184) in the human COL1A2 promoter as the site through which TGF-beta mediates collagen gene expression in human SMC. Gel mobility shift assays demonstrated that this 150 bp DNA fragment formed conjugates with multiple nuclear factors derived from SMC, a process that was further enhanced by TGF-beta.Conclusion. TGF-beta stimulates the human type I collagen gene via a DNA element located in the proximal region of its promoter. Interventions that disrupt interaction between this DNA element and nuclear factors may block the production of collagen in response to TGF-beta and consequently may have a significant effect on the development of intimal hyperplasia. (C) 2003 Elsevier Science (USA).