Home LiteratureArticle Details
PMID: 9033287 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Intestinal epithelial restitution. Involvement of specific laminin isoforms and integrin laminin receptors in wound closure of a transformed model epithelium.

The American journal of pathology ·Vol. 150 ·No. 2 ·1997-02-00 ·Pages 747-60

Lotz MM, Nusrat A, Madara JL, Ezzell R, Wewer UM, Mercurio AM

Abstract

Disruptions in the mucosal lining of the gastrointestinal tract reseal by epithelial cell migration, a process termed restitution. We examined the involvement of laminin isoforms and their integrin receptors in restitution using the intestinal epithelial cell line T84. T84 cells express primarily laminins 5, 6, and 7 as indicated by immunostaining using laminin subunit-specific monoclonal antibodies (MAbs). A MAb (BM2) specific for the laminin alpha 3 subunit, a component of laminins 5, 6, and 7, completely inhibited the closure of mechanical wounds in T84 monolayers. Confocal microscopy using MAbs BM2 (laminin alpha 3 subunit) and 6F12 (laminin beta 3 subunit) revealed that laminin-5 is deposited in a basal matrix that extends into the wound. The MAbs 4E10 (laminin beta 1 subunit) and C4 (laminin beta 2 subunit) stained the lateral membranes between T84 cells. This staining was enhanced in cells adjoining wounds. Because T84 cells stained faintly with MAbs 4C7 (laminin alpha 1 subunit) and with MAbs 4F11 and 1B4 (laminin alpha 2 subunit), we suggest that expression of laminins 6 and 7 is enhanced in response to wounding. The alpha 3 beta 1 integrin and the alpha 6-containing integrins function in wound closure because MAbs specific for the beta 1 integrin subunit (MAb13), the alpha 3 subunit (IVA5), and the alpha 6 subunit (2B7) potently inhibited T84 migration into wounds. Immunofluorescence using UMA9, a beta 4-integrin-specific MAb, revealed that alpha 6 beta 4 integrin exists in a Triton-X-100-insoluble structure at the basal surface and that the staining of this structure is enhanced in cells adjoining wounds. In addition, a Triton-X-100-soluble pool of alpha 6 beta 4, as well as alpha 3 beta 1 and presumably alpha 6 beta 1, was found along lateral surfaces of T84 cells. On flattened cells adjoining wounds, staining for these integrins was distributed diffusely, suggesting a redistribution that accompanies cell migration. Taken together, these data suggest that wound-induced epithelial cell migration is a finely tuned process that is dependent upon the regulated function and localization of specific laminins and their integrin receptors.

MeSH Terms
Antigens, CD/physiology Fluorescent Antibody Technique Humans Integrin alpha3beta1 Integrin alpha6 Integrins/physiology Intestinal Mucosa/physiopathology Isomerism Laminin/physiology Receptors, Laminin/physiology Tumor Cells, Cultured Wound Healing
Chemicals
Antigens, CD Integrin alpha3beta1 Integrin alpha6 Integrins Laminin Receptors, Laminin
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Lotz M M
Division of Gastroenterology, Beth Israel Hospital, Boston, MA 02215, USA.
Nusrat A
Madara J L
Ezzell R
Wewer U M
Mercurio A M
References (64)
64 references, click to expand
  1. The A9 antigen associated with aggressive human squamous carcinoma is structurally and functionally similar to the newly defined integrin alpha 6 beta 4.
    Cancer Res. 1991 May 1;51(9):2395-402 PMID: 1750876
  2. Collagens facilitate epithelial migration in restitution of native guinea pig intestinal epithelium.
    Gastroenterology. 1992 Jan;102(1):119-30 PMID: 1727745
  3. Human laminin isolated in a nearly intact, biologically active form from placenta by limited proteolysis.
    J Biol Chem. 1983 Oct 25;258(20):12654-60 PMID: 6415055
  4. Transforming growth factor beta regulation of migration in wounded rat intestinal epithelial monolayers.
    Gastroenterology. 1993 Jul;105(1):93-101 PMID: 8514065
  5. Human enterocyte (Caco-2) migration is modulated in vitro by extracellular matrix composition and epidermal growth factor.
    J Clin Invest. 1992 Jul;90(1):15-23 PMID: 1634605
  6. Human beta 2 chain of laminin (formerly S chain): cDNA cloning, chromosomal localization, and expression in carcinomas.
    Genomics. 1994 Nov 15;24(2):243-52 PMID: 7698745
  7. Kalinin: an epithelium-specific basement membrane adhesion molecule that is a component of anchoring filaments.
    J Cell Biol. 1991 Aug;114(3):567-76 PMID: 1860885
  8. Redistribution of the hemidesmosome components alpha 6 beta 4 integrin and bullous pemphigoid antigens during epithelial wound healing.
    Exp Cell Res. 1993 Jul;207(1):86-98 PMID: 8319775
  9. Epithelial-mesenchymal interactions in the production of basement membrane components in the gut.
    Development. 1988 Feb;102(2):339-47 PMID: 17061377
  10. The gamma 2 chain of kalinin/laminin 5 is preferentially expressed in invading malignant cells in human cancers.
    Am J Pathol. 1994 Oct;145(4):782-91 PMID: 7943170
  11. Fibronectin and wound healing.
    J Cell Biochem. 1984;26(2):107-16 PMID: 6084665
  12. Intestinal epithelial restitution. Characterization of a cell culture model and mapping of cytoskeletal elements in migrating cells.
    J Clin Invest. 1992 May;89(5):1501-11 PMID: 1569187
  13. Ca(2+)- and calcineurin-dependent recycling of an integrin to the front of migrating neutrophils.
    Nature. 1995 Sep 7;377(6544):75-9 PMID: 7544874
  14. Mutations in the gamma 2 chain gene (LAMC2) of kalinin/laminin 5 in the junctional forms of epidermolysis bullosa.
    Nat Genet. 1994 Mar;6(3):293-7 PMID: 8012393
  15. Regulation of fibronectin biosynthesis by dexamethasone, transforming growth factor beta, and cAMP in human cell lines.
    J Cell Biol. 1988 Jun;106(6):2159-70 PMID: 2454932
  16. Marked stimulation of cell adhesion and motility by ladsin, a laminin-like scatter factor.
    J Biochem. 1994 Oct;116(4):862-9 PMID: 7533763
  17. Surface relocation of alpha 6 beta 4 integrins and assembly of hemidesmosomes in an in vitro model of wound healing.
    J Cell Biol. 1991 Dec;115(6):1737-50 PMID: 1757471
  18. Inhibition of laminin alpha 1-chain expression leads to alteration of basement membrane assembly and cell differentiation.
    J Cell Biol. 1996 Apr;133(2):417-30 PMID: 8609173
  19. Integrin alpha 6/beta 4 complex is located in hemidesmosomes, suggesting a major role in epidermal cell-basement membrane adhesion.
    J Cell Biol. 1991 May;113(4):907-17 PMID: 2026654
  20. Integrin-cytoskeletal interactions in migrating fibroblasts are dynamic, asymmetric, and regulated.
    J Cell Biol. 1993 Nov;123(4):977-91 PMID: 8227153
  21. Independent modulation of enterocyte migration and proliferation by growth factors, matrix proteins, and pharmacologic agents in an in vitro model of mucosal healing.
    Surgery. 1992 Aug;112(2):299-307; discussion 307-8 PMID: 1353641
  22. Analysis of fibronectin receptor function with monoclonal antibodies: roles in cell adhesion, migration, matrix assembly, and cytoskeletal organization.
    J Cell Biol. 1989 Aug;109(2):863-75 PMID: 2527241
  23. Reciprocal expression of laminin A-chain isoforms along the crypt-villus axis in the human small intestine.
    Gastroenterology. 1994 Apr;106(4):829-39 PMID: 8143989
  24. Established intestinal cell lines as model systems for electrolyte transport studies.
    Methods Enzymol. 1990;192:354-89 PMID: 2074798
  25. Self-assembly and calcium-binding sites in laminin. A three-arm interaction model.
    J Biol Chem. 1993 Aug 15;268(23):17286-99 PMID: 8349613
  26. Distribution and isolation of four laminin variants; tissue restricted distribution of heterotrimers assembled from five different subunits.
    Cell Regul. 1990 Sep;1(10):731-40 PMID: 2099832
  27. Laminin receptors: achieving specificity through cooperation.
    Trends Cell Biol. 1995 Nov;5(11):419-23 PMID: 14732046
  28. Hepatocyte growth factor/scatter factor effects on epithelia. Regulation of intercellular junctions in transformed and nontransformed cell lines, basolateral polarization of c-met receptor in transformed and natural intestinal epithelia, and induction of rapid wound repair in a transformed model epithelium.
    J Clin Invest. 1994 May;93(5):2056-65 PMID: 8182137
  29. Laminin mediates tissue-specific gene expression in mammary epithelia.
    J Cell Biol. 1995 May;129(3):591-603 PMID: 7730398
  30. Distinctive integrin expression in the newly forming epidermis during wound healing in humans.
    J Invest Dermatol. 1993 Oct;101(4):600-4 PMID: 8409530
  31. Merosin, a protein specific for basement membranes of Schwann cells, striated muscle, and trophoblast, is expressed late in nerve and muscle development.
    Proc Natl Acad Sci U S A. 1988 Mar;85(5):1544-8 PMID: 3278318
  32. Hemidesmosomes: extracellular matrix/intermediate filament connectors.
    Exp Cell Res. 1994 Jul;213(1):1-11 PMID: 8020577
  33. Repair of rabbit duodenal mucosa after acid injury in vivo and in vitro.
    Gastroenterology. 1987 Jun;92(6):1973-86 PMID: 3569771
  34. Developmental expression of laminin-5 and HD1 in the intestine: epithelial to mesenchymal shift for the laminin gamma-2 chain subunit deposition.
    Dev Dyn. 1996 May;206(1):12-23 PMID: 9019243
  35. Dual and asynchronous deposition of laminin chains at the epithelial-mesenchymal interface in the gut.
    Gastroenterology. 1992 Jun;102(6):1835-45 PMID: 1587403
  36. Changes in the expression of laminin during intestinal development.
    Development. 1991 Jun;112(2):477-87 PMID: 1794317
  37. Role of laminin A chain in the development of epithelial cell polarity.
    Cell. 1988 Oct 21;55(2):331-41 PMID: 3048705
  38. Mapping of domains in human laminin using monoclonal antibodies: localization of the neurite-promoting site.
    J Cell Biol. 1986 Dec;103(6 Pt 1):2457-65 PMID: 2430984
  39. Hepatocyte growth factor/scatter factor modulates intestinal epithelial cell proliferation and migration.
    Biochem Biophys Res Commun. 1994 Jul 29;202(2):701-9 PMID: 8048940
  40. Molecular heterogeneity of basal laminae: isoforms of laminin and collagen IV at the neuromuscular junction and elsewhere.
    J Cell Biol. 1990 Oct;111(4):1685-99 PMID: 2211832
  41. Fibroblast growth factors modulate intestinal epithelial cell growth and migration.
    Gastroenterology. 1994 May;106(5):1254-62 PMID: 7513666
  42. The anchoring filament protein kalinin is synthesized and secreted as a high molecular weight precursor.
    J Biol Chem. 1992 Sep 5;267(25):17900-6 PMID: 1517226
  43. Inside-out integrin signaling in macrophages. Analysis of the role of the alpha 6A beta 1 and alpha 6B beta 1 integrin variants in laminin adhesion by cDNA expression in an alpha 6 integrin-deficient macrophage cell line.
    J Biol Chem. 1993 May 25;268(15):11401-8 PMID: 8496190
  44. Alpha 6 beta 4 integrin heterodimer is a component of hemidesmosomes.
    Proc Natl Acad Sci U S A. 1990 Nov;87(22):8970-4 PMID: 2247472
  45. Morphology and electrophysiology of guinea pig gastric mucosal repair in vitro.
    Am J Physiol. 1983 Feb;244(2):G171-82 PMID: 6824088
  46. Restitution of frog gastric mucosa in vitro: effect of basic fibroblast growth factor.
    Gastroenterology. 1993 May;104(5):1337-45 PMID: 8482448
  47. Differential expression of laminin isoforms and alpha 6-beta 4 integrin subunits in the developing human and mouse intestine.
    Dev Dyn. 1994 Sep;201(1):71-85 PMID: 7803849
  48. Wound repair in the context of extracellular matrix.
    Curr Opin Cell Biol. 1994 Oct;6(5):717-25 PMID: 7530463
  49. Origin and deposition of basement membrane heparan sulfate proteoglycan in the developing intestine.
    J Cell Biol. 1989 Oct;109(4 Pt 1):1837-48 PMID: 2529263
  50. Fibronectin and fibrin provide a provisional matrix for epidermal cell migration during wound reepithelialization.
    J Invest Dermatol. 1982 Nov;79(5):264-9 PMID: 6752288
  51. Human amnion contains a novel laminin variant, laminin 7, which like laminin 6, covalently associates with laminin 5 to promote stable epithelial-stromal attachment.
    J Cell Biol. 1996 Mar;132(6):1189-98 PMID: 8601594
  52. Intestinal epithelia (T84) possess basolateral ligands for CD11b/CD18-mediated neutrophil adherence.
    Am J Physiol. 1995 Feb;268(2 Pt 1):C472-9 PMID: 7532360
  53. Rapid epithelial restitution of the rat gastric mucosa after ethanol injury.
    Lab Invest. 1984 Nov;51(5):573-83 PMID: 6492759
  54. Transforming growth factor-beta switches the pattern of integrins expressed in MG-63 human osteosarcoma cells and causes a selective loss of cell adhesion to laminin.
    J Biol Chem. 1989 Dec 25;264(36):21806-11 PMID: 2600089
  55. Role of laminin in the attachment of PAM 212 (epithelial) cells to basement membrane collagen.
    Cell. 1980 Dec;22(3):719-26 PMID: 7460011
  56. Transforming growth factor-beta 1 modulates beta 1 and beta 5 integrin receptors and induces the de novo expression of the alpha v beta 6 heterodimer in normal human keratinocytes: implications for wound healing.
    J Cell Biol. 1995 May;129(3):853-65 PMID: 7537276
  57. Domains of laminin with growth-factor activity.
    Cell. 1989 Jan 13;56(1):93-101 PMID: 2491959
  58. Rapid barrier restitution in an in vitro model of intestinal epithelial injury.
    Lab Invest. 1989 Feb;60(2):237-44 PMID: 2915518
  59. Growth factor-induced acceleration of tissue repair through direct and inductive activities in a rabbit dermal ulcer model.
    J Clin Invest. 1991 Feb;87(2):694-703 PMID: 1991853
  60. Biological activities of laminin.
    J Cell Biochem. 1985;27(4):317-25 PMID: 3889019
  61. A recombinant tail-less integrin beta 4 subunit disrupts hemidesmosomes, but does not suppress alpha 6 beta 4-mediated cell adhesion to laminins.
    J Cell Biol. 1995 Apr;129(2):473-87 PMID: 7721947
  62. Restitution of the surface epithelium of the in vitro frog gastric mucosa after damage with hyperosmolar sodium chloride. Morphologic and physiologic characteristics.
    Gastroenterology. 1982 Jun;82(6):1409-26 PMID: 6978275
  63. Cytokine modulation of intestinal epithelial cell restitution: central role of transforming growth factor beta.
    Gastroenterology. 1993 Nov;105(5):1323-32 PMID: 8224636
  64. Regulation of extracellular matrix proteins and integrin cell substratum adhesion receptors on epithelium during cutaneous human wound healing in vivo.
    Am J Pathol. 1993 Nov;143(5):1458-69 PMID: 7694470
Article Info
Journal
The American journal of pathology
Abbr.
Am J Pathol
ISSN
0002-9440
Published
1997-02-00
Pages
747-60
Language
English
Region
United States
NLM ID
0370502
PMCID
PMC1858283
Subset
IM
Grants
NCI NIH HHS · CA42274 · United States
NIDDK NIH HHS · DK02130 · United States
NIDDK NIH HHS · DK37932 · United States
External Links
PubMed source PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2