Identifying the Regulative Role of NF-κB Binding Sites Within Promoter Region of Human Matrix Metalloproteinase 9 (mmp-9) by TNF-α Induction
Identifying the Regulative Role of NF-κB Binding Sites Within Promoter Region of Human Matrix Metalloproteinase 9 (mmp-9) by TNF-α Induction
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DOI：
10.1007/s12010-012-9958-3
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发表时间：
2013-01-01
影响因子：
3
通讯作者：
Lin, Chih Sheng
中科院分区：
文献类型：
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作者：
Wu, Hsi Tien;Sie, Syu Sheng;Lin, Chih Sheng
基质金属蛋白酶9（MMP-9）是MMP家族的成员之一，参与多种生理过程，包括心血管疾病（CVD）。肿瘤坏死因子-α（TNF-α）被认为是具有多效性生物学能力的细胞因子，并且当TNF-α异常释放并刺激MMP-9表达和活化时导致CVD过程。在这项研究中，我们研究了TNF-α调节MMP-9表达的分子机制。实验结果证实TNF-α可上调大鼠心肌成肌细胞H9 c2中MMP-9的表达。为了从转录水平研究MMP-9的调控作用，克隆了人MMP-9启动子区的2.2kb（-2168/+18）片段，并将其构建到荧光素酶报告基因载体中。2.2-kb序列被鉴定为具有三个候选核因子-κ B（NF-κ B）结合位点：NF-κ B I（-1418/-1409）、NF-κ B II（-626/-617）和NF-κ B III（-353/-345）。构建了一系列MMP-9启动子NF-κ B B位点突变的报告载体，并转染H9 c2细胞。结果表明，MMP-9启动子区域内的NF-κ B II结合位点（-626/-617）在TNF-α诱导的MMP-9表达上调中起关键作用。此外，我们还首次发现NF-κ B I与c-Rel相似，可能是调节MMP-9表达的NF-κ B家族之一。
Matrix metalloproteinase 9 (MMP-9), a member of MMP family, is involved in many physiological processes, including cardiovascular disease (CVD). Tumor necrosis factor-alpha (TNF-alpha) is considered a cytokine with pleiotropic biological capabilities and leads to the process of CVD when TNF-alpha is abnormally released and stimulates MMP-9 expression and activation. In this study, we investigated the molecular mechanism of TNF-alpha-regulated MMP-9 expression. The experimental results confirm that TNF-alpha could upregulate MMP-9 expression in heart myoblast H9c2 cells of rat. To evaluate the MMP-9 regulation at transcriptional level, a DNA fragment of 2.2 kb (-2168/+18) of human mmp-9 promoter region was cloned and constructed in a vector of luciferase reporter gene. The 2.2-kb sequences were identified as having three candidate nuclear factor-kappa B (NF-kappa B) binding sites: NF-kappa B I (-1418/-1409), NF-kappa B II (-626/-617), and NF-kappa B III (-353/-345). A series of reporter vectors with the mutated NF-kappa B sites of mmp-9 promoter sequences were constructed and transfected into H9c2 cells. The results show that the NF-kappa B II binding site (-626/-617) within the promoter region of mmp-9 plays a key role in upregulation of mmp-9 expression by TNF-alpha induction. In addition, we also first identified that the NF-kappa B I, similar to c-Rel, might be one of the NF-kappa B families to regulate mmp-9 expression.