title: ADAM-15 inhibits wound healing in human intestinal epithelial cell monolayers.
abstract: 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.
authors.affiliation: Department of Medicine, Division of Digestive Diseases, Emory University School of Medicine, Atlanta, GA 30322, USA.
authors.forename: Laetitia
authors.initials: L
authors.lastname: Charrier
authors.forename: Yutao
authors.initials: Y
authors.lastname: Yan
authors.forename: Adel
authors.initials: A
authors.lastname: Driss
authors.forename: Christian L
authors.initials: CL
authors.lastname: Laboisse
authors.forename: Shanthi V
authors.initials: SV
authors.lastname: Sitaraman
authors.forename: Didier
authors.initials: D
authors.lastname: Merlin
mesh_terms: D051722:ADAM Proteins; D000072197:ADAM10 Protein; D000072199:ADAM12 Protein; D000595:Amino Acid Sequence; D053829:Amyloid Precursor Protein Secretases; D018938:Caco-2 Cells; D002465:Cell Movement / Q000502:physiology; D017097:Electric Impedance; D015870:Gene Expression; D019073:HT29 Cells; D006801:Humans; D007413:Intestinal Mucosa / Q000737:chemistry / Q000166:cytology / Q000502:physiology*; D016015:Logistic Models; D008565:Membrane Proteins / Q000032:analysis / Q000737:chemistry / Q000502:physiology*; D008666:Metalloendopeptidases / Q000032:analysis / Q000737:chemistry / Q000502:physiology*; D008969:Molecular Sequence Data; D012333:RNA, Messenger; D014945:Wound Healing / Q000502:physiology
chemical_list: D008565:Membrane Proteins; D012333:RNA, Messenger; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D000072199:ADAM12 Protein; C000606480:ADAM12 protein, human; C098660:ADAM15 protein, human; D008666:Metalloendopeptidases; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human