Induction of MMP-1 (collagenase-1) by relaxin in fibrocartilaginous cells requires both the AP-1 and PEA-3 promoter sites.
Induction of MMP-1 (collagenase-1) by relaxin in fibrocartilaginous cells requires both the AP-1 and PEA-3 promoter sites.
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DOI：
10.1111/j.1601-6343.2009.01451.x
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发表时间：
2009-08
影响因子：
3.1
通讯作者：
Wang W
中科院分区：
文献类型：
--
作者：
Kapila S;Xie Y;Wang W
松弛素诱导TMJ纤维软骨细胞中的基质金属蛋白酶MMP-1（胶原酶-1），β-雌二醇可增强这种反应。我们确定了MMP-1的启动子位点和转录因子，松弛素诱导或不诱导β-雌二醇在纤维软骨细胞。用含有调节氯霉素酰基转移酶（CAT）基因的人MMP-1启动子的特定片段的pBLCAT 2质粒瞬时转染早期传代细胞，并与含有β-半乳糖苷酶基因的质粒共转染。将细胞在单独的无血清培养基或含有0.1 ng/ml松弛素或20 ng/ml β-雌二醇或两种激素的培养基中培养，并测定裂解物的CAT和β-半乳糖苷酶活性。转染了-1200/-42或-139/-42 bp MMP-1启动子-报告基因构建体的细胞，在β-雌二醇存在或不存在的情况下，松弛素对CAT的诱导分别为1.5倍和3倍。松弛素在MMP-1启动子的−137/−69区域（包含AP-1和PEA 3结合位点）缺失时不能诱导CAT。使用野生型或突变的最小AP-1和PEA-3启动子，我们发现，这两个启动子位点是必不可少的诱导MMP-1松弛素。转录因子c-fos和c-jun（共同形成AP-1异源二聚体）以及调节PEA-3位点的Ets-1的mRNA被松弛素或β-雌二醇加松弛素上调。这些研究表明，AP-1和PEA-3启动子位点都是纤维软骨细胞中松弛素诱导MMP-1所必需的。
Relaxin induces the matrix metalloproteinase MMP-1 (collagenase-1) in TMJ fibrocartilaginous cells, and this response is potentiated by β-estradiol. We identified the MMP-1 promoter sites and transcription factors that are induced by relaxin with or without β-estradiol in fibrocartilaginous cells. Early passage cells were transiently transfected with the pBLCAT2 plasmid containing specific segments of the human MMP-1 promoter regulating the chloramphenicol acyl transferase (CAT) gene and co-transfected with a plasmid containing the β-galactosidase gene. The cells were cultured in serum-free medium alone or medium containing 0.1 ng/ml relaxin, or 20 ng/ml β-estradiol or both hormones, and lysates assayed for CAT and β-galactosidase activity. Cells transfected with the −1200/−42 or −139/−42 bp MMP-1 promoter-reporter constructs showed 1.5-fold and 3-fold induction of CAT by relaxin in the absence or presence of β-estradiol, respectively. Relaxin failed to induce CAT in the absence of the −137/−69 region of the MMP-1 promoter, which contains the AP-1- and PEA3-binding sites. Using wild type or mutated minimal AP-1 and PEA-3 promoters we found that both these promoter sites are essential for the induction of MMP-1 by relaxin. The mRNAs for transcription factors c-fos and c-jun, which together form the AP-1 heterodimer, and Ets-1 that modulates the PEA-3 site, were upregulated by relaxin or β-estradiol plus relaxin. These studies show that both the AP-1 and PEA-3 promoter sites are necessary for the induction of MMP-1 by relaxin in fibrocartilaginous cells.
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影响因子：
2.9
作者：
AUBLE, DT;BRINCKERHOFF, CE
通讯作者：
BRINCKERHOFF, CE
影响因子：
3.9
作者：
O'Brien, RM;Streeper, RS;Hornbuckle, LA
通讯作者：
Hornbuckle, LA
影响因子：
4.1
作者：
Anand-Ivell, Ravinder;Heng, Kee;Ivell, Richard
通讯作者：
Ivell, Richard
影响因子：
4.8
作者：
Ayala, JE;Streeper, RS;O'Brien, RM
通讯作者：
O'Brien, RM
影响因子：
--
作者：
Hallberg, LM;Ikeno, Y;Greeley, GH
通讯作者：
Greeley, GH