Mercer Becky A, Wallace Alison M, Brinckerhoff Constance E, D'Armiento Jeanine M
Department of Medicine, Division of Molecular and Pulmonary Medicine, College of Physicians and Surgeons, Columbia University, 630 West 168th Street, New York, NY 10032, USA.
Am J Respir Cell Mol Biol. 2009 Jan;40(1):4-12. doi: 10.1165/rcmb.2007-0310OC. Epub 2008 Jul 10.
Tobacco-related diseases are leading causes of death worldwide, and many are associated with expression of matrix metalloproteinase-1 (MMP-1). We have reported extracellular signal-regulated kinase (ERK)1/2-dependent induction of MMP-1 by cigarette smoke in lung epithelial cells. Our objectives were to define regions of the human MMP-1 promoter required for activation by smoke, to identify differences in responses of the 1G/2G -1607 polymorphic promoters to smoke, and to identify relevant transcription factors whose activity in airway epithelial cells is increased by smoke. The responses of deletion and mutant promoter constructs were measured in transfected cells during exposure to cigarette smoke extract (CSE). DNA oligonucleotide arrays were used to identify transcription factors activated after smoke exposure. CSE activated the MMP-1 promoter, and this induction was prevented by PD98059 blockade of ERK1/2 phosphorylation. Deletion studies revealed the distal 1kb promoter region (-4438 to -3280 upstream of the transcription start site) is essential for CSE induction of MMP-1, and confers activation of a minimal promoter. Studies of 1G and 2G MMP-1 polymorphic promoter variants revealed higher 2G allele basal and CSE-responsive activities than the 1G allele. Cotransfection, mithramycin, and electrophoretic mobility shift assay studies identified activating and repressive roles for Sp1 and PEA3 transcription factors, respectively. Oligonucleotide DNA arrays confirmed activation of Sp1 and PEA3 by CSE. These data demonstrate that the MMP-1 promoter is a direct target of cigarette smoke in lung epithelial cells. This characterization of a smoke response region in the distal MMP-1 promoter has implications for smoking-related diseases such as cancer, heart disease, and emphysema.
烟草相关疾病是全球主要死因，其中许多与基质金属蛋白酶-1（MMP-1）的表达有关。我们曾报道香烟烟雾在肺上皮细胞中通过细胞外信号调节激酶（ERK）1/2依赖性诱导MMP-1。我们的目标是确定烟雾激活人MMP-1启动子所需的区域，识别1G/2G -1607多态性启动子对烟雾反应的差异，并确定气道上皮细胞中活性因烟雾而增加的相关转录因子。在转染细胞暴露于香烟烟雾提取物（CSE）期间，测量缺失和突变启动子构建体的反应。使用DNA寡核苷酸阵列鉴定烟雾暴露后激活的转录因子。CSE激活了MMP-1启动子，而ERK1/2磷酸化的PD98059阻断可阻止这种诱导。缺失研究表明，远端1kb启动子区域（转录起始位点上游-4438至-3280）对于CSE诱导MMP-1至关重要，并赋予最小启动子激活。对1G和2G MMP-1多态性启动子变体的研究表明，2G等位基因的基础活性和CSE反应活性高于1G等位基因。共转染、光神霉素和电泳迁移率变动分析研究分别确定了Sp1和PEA3转录因子的激活和抑制作用。寡核苷酸DNA阵列证实CSE激活了Sp1和PEA3。这些数据表明，MMP-1启动子是肺上皮细胞中香烟烟雾的直接靶点。MMP-1启动子远端烟雾反应区域的这一特征对癌症、心脏病和肺气肿等吸烟相关疾病具有重要意义。
