ADAM-15 inhibits wound healing in human intestinal epithelial cell monolayers
ADAM-15 inhibits wound healing in human intestinal epithelial cell monolayers
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DOI：
10.1152/ajpgi.00262.2004
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发表时间：
2005-02-01
影响因子：
4.5
通讯作者：
Merlin, D
中科院分区：
文献类型：
--
作者：
Charrier, L;Yan, YT;Merlin, D
去整合素金属蛋白酶（或亚当斯）是膜锚定的糖蛋白，其涉及细胞-细胞或细胞-基质相互作用以及细胞表面上分子的蛋白水解。肠上皮细胞中亚当斯的表达和/或病理生理学意义尚不清楚。因此，我们的目的是研究亚当斯在肠上皮细胞中的表达及其作用。通过RT-PCR、Western blot和免疫荧光实验检测亚当斯的表达。利用电电池基底阻抗传感技术进行伤口愈合实验。我们的研究结果表明，亚当斯-10，-12，和-15 mRNA在结肠人细胞系Caco 2-BBE和HT 29-Cl. 19 A中表达。从Caco 2-BBE poly（A）（+）RNA克隆并包含整个编码区的ADAM-15互补DNA被发现与数据库中保存的ADAM-15序列相比更短，并且存在编码胞质尾区的不同区域。在Caco 2-BBE细胞和结肠上皮细胞中，发现ADAM-15蛋白存在于顶端、基底外侧和细胞内室。我们还表明，在Caco 2-BBE单层的伤口愈合试验中，ADAM-15的过表达减少了细胞迁移。我们的数据显示，1）ADAM-15在人肠上皮细胞中表达，2）ADAM-15的一种新变体在人肠上皮细胞系中表达，3）ADAM-15参与肠上皮细胞伤口愈合过程。总之，这些结果表明，ADAM-15可能在肠细胞中具有重要的病理生理作用。
The disintegrin metalloproteases (or ADAMs) are membrane-anchored glycoproteins that have been implicated in cell-cell or cell-matrix interactions and in proteolysis of molecules on the cell surface. The expression and/or the pathophysiological implications of ADAMs are not known in intestinal epithelial cells. Therefore, our aim was to investigate the expression and the role of ADAMs in intestinal epithelial cells. Expression of ADAMs was assessed by RT-PCR, Western blot analysis, and immunufluorescence experiments. Wound-healing experiments were performed by using the electric cell substrate impedence sensing technology. Our results showed that ADAMs-10, -12, and -15 mRNA are expressed in the colonic human cell lines Caco2-BBE and HT29-Cl.19A. An ADAM-15 complementary DNA cloned from Caco2-BBE poly(A)(+) RNA, and encompassing the entire coding region, was found to be shorter and to present a different region encoding the cytoplasmic tail compared with ADAM-15 sequence deposited in the database. In Caco2-BBE cells and colonic epithelial cells, ADAM-15 protein was found in the apical, basolateral, and intracellular compartments. We also showed that the overexpression of ADAM-15 reduced cell migration in a wound-healing assay in Caco2-BBE monolayers. Our data show that 1) ADAM-15 is expressed in human intestinal epithelia, 2) a new variant of ADAM-15 is expressed in a human intestinal epithelial cell line, and 3) ADAM-15 is involved in intestinal epithelial cells wound-healing processes. Together, these results suggest that ADAM-15 may have important pathophysiological roles in intestinal cells.