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Exogenous phospholipase C stimulates epithelial cell migration and integrin expression in vitro
J D Firth1, E E Putnins, H Larjava
1Department of Oral Biological and Medical Sciences, University of British Columbia, Vancouver, British Columbia, Canada.
Summary
Bacterial phospholipase C disrupts epithelial healing by increasing cell migration and altering integrin expression. This bacterial enzyme impedes wound repair by affecting cell-cell contact and extracellular matrix interactions.
Area of Science:
- Cell Biology
- Microbiology
- Dermatology
Background:
- Bacterial phospholipase C (PLC) is a virulence factor in human diseases.
- PLC impedes wound healing by affecting epithelial cell behavior.
Purpose of the Study:
- To investigate the role of bacterial PLC in epithelial cell growth and migration.
- To understand the signaling pathways and molecular mechanisms involved in PLC-induced epithelial disruption.
Main Methods:
- Cultured human skin keratinocytes under simulated wound healing conditions.
- Utilized inhibitors of phosphoinositol signaling (neomycin sulfate) and protein kinase C (RO-31-8220).
- Assessed matrix metalloproteinase-9 (MMP-9) levels and integrin expression (alpha(v), alpha5, beta1) via immunostaining and confocal microscopy.
Main Results:
- Bacterial PLC decreased cell-cell contact and increased keratinocyte migration, disrupting epithelial sheets.
- PLC-induced migration was inhibited by neomycin sulfate and RO-31-8220.
- Elevated MMP-9 levels and enhanced cell adhesion to fibronectin, vitronectin, and collagen IV were observed.
- Increased expression and basal deposition of integrin subunits beta1 in focal contacts were noted.
Conclusions:
- Exogenous bacterial PLC perturbs normal reepithelialization.
- Altered integrin expression and MMP-9 activity are key mechanisms in PLC-induced disruption of wound healing.