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ADAM-15 inhibits wound healing in human intestinal epithelial cell monolayers
Laetitia Charrier1, Yutao Yan, Adel Driss
1Department of Medicine, Division of Digestive Diseases, Emory University School of Medicine, Atlanta, GA 30322, USA.
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.
Insights
Disintegrin metalloproteases (ADAMs) like ADAM-15 are expressed in human intestinal cells and influence cell migration during wound healing, suggesting a role in intestinal health.
Area of Science:
- Cell Biology
- Molecular Biology
- Gastroenterology
Background:
- Disintegrin metalloproteases (ADAMs) are crucial for cell interactions and surface protein regulation.
- The role and expression of ADAMs in intestinal epithelial cells remain largely unknown.
- Investigating ADAMs in the gut is vital for understanding intestinal epithelial cell function.
Purpose of the Study:
- To determine the expression of ADAMs in human intestinal epithelial cells.
- To elucidate the function of ADAM-15 in intestinal epithelial cell migration and wound healing.
Main Methods:
- Gene expression analysis using RT-PCR.
- Protein expression validation via Western blot and immunofluorescence.
- Cell migration assessment using electric cell-substrate impedance sensing (ECIS) for wound-healing assays.
Main Results:
- ADAM-10, ADAM-12, and ADAM-15 mRNA were detected in human colonic cell lines (Caco2-BBE, HT29-Cl.19A).
- A novel ADAM-15 variant with a distinct cytoplasmic tail was identified in Caco2-BBE cells.
- ADAM-15 protein localizes to apical, basolateral, and intracellular compartments in intestinal cells.
- ADAM-15 overexpression significantly reduced cell migration in Caco2-BBE monolayers during wound healing.
Conclusions:
- ADAM-15 is expressed in human intestinal epithelia.
- A new ADAM-15 variant is present in human intestinal epithelial cells.
- ADAM-15 plays a role in the wound-healing process of intestinal epithelial cells.
- ADAM-15 may have significant pathophysiological implications in intestinal diseases.
