Effect of transforming growth factor-beta on activity of connective tissue growth factor gene promoter in mouse NIH/3T3 fibroblasts

• 来源期刊: 中国药理学报（英文版）
• 学科: 医学
• 关键词: transforming growth factor-beta connective tissue growth factor promoter mitogen-activated protein kinases signal transduction
• 年，卷（期）: 2004,（4）
• 页码范围: 485-489
• 页数: 5页
• 分类号: R9
• 语种: 英文

摘要

AIM: To investigate the regulatory mechanism of transforming growth factor-beta on activity of connective tissue growth factor promoter in mouse NIH/3T3 fibroblasts. METHODS: The regulation fragment of the 5' flanking region of the human CTGF gene was linked to pGL3-Basic vector, a firefly luciferase reporter construct without promoter. The recombinant plasmid pCTGF-luc was transiently transfected to NIH/3T3 fibroblasts. The activity of CTGF promoter after treatment of TGF-β1 and MAPK pathway inhibitors were assayed with luciferase reporter gene assay system. RESULTS: TGF-β1-induced increase of CTGF promoter activity was concentration-dependent,with a plateau at 5 μg/L by 2.67-fold vs control (P<0.05). The TGF-β1 stimulation of CTGF promoter activity was time-dependent, too. After exposure to TGF-β1 (5 μg/L), the maximal level of luciferase activity was reached at 12 h and maintained to 24 h by 2.76- and 2.20-fold vs control, respectively (P<0.05). Blockade of mitogen-activated protein kinases (MAPK) pathway with PD98059 (10 μmnol/L), the MAP kinase kinase 1 inhibitor, and SB203580 (10μmol/L), the p38 MAP kinase inhibitor, decreased basal and TGF-β1-induced activation of CTGF promoter. However,inhibition of c-Jun-N-terminal kinase/stress-activated protein kinase by SP600125 (20 μmol/L) was without effect.CONCLUSION: TGF-β1 stimulated the transcriptional activity of CTGF gene promoter in NIH/3T3 fibroblasts in a dose- and time-dependent manner. MAPK pathway may play a role in the regulation of TGF-β1-induced CTGF expression.