The alpha3 laminin subunit, alpha6beta4 and alpha3beta1 integrin coordinately regulate wound healing in cultured epithelial cells and in the skin.
Previously, we demonstrated that proteolytic processing within the globular domain of the alpha3 subunit of laminin-5 (LN5) converts LN5 from a cell motility-inducing factor to a protein complex that can trigger the formation of hemidesmosomes, certain cell-matrix attachment sites found in epithelial cells. We have prepared a monoclonal antibody (12C4) whose epitope is located toward the carboxy terminus of the globular domain of the alpha3 laminin subunit. This epitope is lost from the alpha3 subunit as a consequence of proteolytic processing. Antibody 12C4 stains throughout the matrix of cells that fail to process the alpha3 laminin subunit, but does not recognize the matrix of confluent cultures of MCF-10A cells, which efficiently process their alpha3 laminin chain. In subconfluent populations of MCF-10A cells, 12C4 only stains matrix deposited at the outer edges of cell colonies. In these cells, integrin alpha3beta1 occasionally colocalizes with the staining generated by the 12C4 antibody but alpha6beta4 integrin does not. In wounded MCF-10A cell cultures, the 12C4 antibody stains the extracellular matrix beneath those cells at the very edge of the cellular sheet that moves to cover the wound site. A similar phenomenon is observed in human skin wounds, since we also detect expression of the unprocessed alpha3 laminin subunit at the leading tip of the sheet of epidermal cells that epithelializes skin wounds in vivo. In addition, using alpha3 laminin subunit and integrin function-inhibiting antibodies, we provide evidence that LN5 and its two integrin receptors (alpha6beta4 and alpha3beta1) appear necessary for wound healing to occur in MCF-10A cell culture wounds. We propose a model for healing of wounded epithelial tissues based on these results.
Department of Cell and Molecular Biology, Northwestern University Medical School, Chicago, IL 60611, USA.
L E
LE
Goldfinger
S B
SB
Hopkinson
G W
GW
deHart
S
S
Collawn
J R
JR
Couchman
J C
JC
Jones
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D000954:Antigens, Surface / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D001943:Breast Neoplasms; D002448:Cell Adhesion / Q000502:physiology; D002465:Cell Movement / Q000502:physiology; D003896:Desmosomes / Q000502:physiology; D004847:Epithelial Cells / Q000737:chemistry / Q000166:cytology* / Q000648:ultrastructure; D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D039201:Integrin alpha3beta1; D039161:Integrin alpha6beta4; D016023:Integrins / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D008854:Microscopy, Electron; D011554:Pseudopodia / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D017474:Receptors, Laminin / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D012867:Skin / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D014407:Tumor Cells, Cultured; D015854:Up-Regulation / Q000502:physiology; D014945:Wound Healing / Q000502:physiology*
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D039201:Integrin alpha3beta1; D039161:Integrin alpha6beta4; D016023:Integrins; D017474:Receptors, Laminin