Thymosin beta4 promotes human conjunctival epithelial cell migration.
OBJECTIVE
In this study the effects of thymosin beta4 (Tbeta4) on migration and production of laminin-5 in the human conjunctival cell line HC0597 was analyzed.


METHODS
Boyden chamber assays assessed the ability of Tbeta4 to stimulate in vitro cell migration. Control or Tbeta4-treated cells were processed for immunofluorescence microscopy using antibodies to vinculin or laminin-5. Cell lysates were processed for Western blot and densitometric analysis using antibodies to laminin-5 alpha3 or gamma2 chains.


RESULTS
Tbeta4 stimulated migration in a dose-dependent manner. Focal adhesions present in Tbeta4-treated cells were smaller and more rounded compared to the "streaks" characteristic of controls. Western blot analysis and densitometry revealed that Tbeta4-treated cells expressed more laminin-5 alpha3 and gamma2 chain protein.


CONCLUSIONS
Tbeta4 stimulates in vitro conjunctival epithelial cell migration, and results in altered focal adhesion formation and increased extracellular laminin-5 deposition. The increased migration may be correlated with increased production of laminin-5.
D015153:Blotting, Western; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D003228:Conjunctiva / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D003720:Densitometry; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000187:drug effects / Q000502:physiology; D005455:Fluorescent Antibody Technique; D006801:Humans; D008856:Microscopy, Fluorescence; D012016:Reference Values; D013947:Thymosin / Q000008:administration & dosage / Q000494:pharmacology*; D000099201:Thymosin beta(4)
D013947:Thymosin; D000099201:Thymosin beta(4)