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Localization of beta 1, beta 3, alpha 5, alpha v, and alpha IIb subunits of
the integrin family in spreading human erythroleukemia cells
J Ylanne, DA Cheresh and I Virtanen
Department of Anatomy, University of Helsinki, Finland.
The localization of five integrin subunit proteins was studied in human
erythroleukemia (HEL) cells spreading on various culture substrata in the
presence of 12-O-tetradecanoylphorbol-13-acetate (TPA) and the absence of
serum. The cells readily adhered on fibronectin, but TPA was needed for
adherence on vitronectin and for the spreading of the cells on both
substrata. Indirect immunofluorescence microscopy showed that in the spread
cells cultured on vitronectin or fibronectin for 2 hours, beta 1, beta 3,
alpha 5, and alpha IIb integrin subunits were localized at focal adhesions
as identified by talin-immunoreactivity. The alpha v integrin
immunoreactivity was initially found at the focal adhesions when the cells
were cultured on vitronectin, but was also found later in cells cultured on
fibronectin. The alpha IIb integrin immunoreactivity disappeared from focal
adhesions within 24 hours. The alpha 5 and beta 1 integrin
immunoreactivities disappeared from the focal adhesions in cells cultured
on vitronectin, but not in cells cultured on fibronectin. When the cells
were plated on glass substratum in the presence of TPA, they spread much
slower than on vitronectin or fibronectin, but some cells showed focal
adhesions after only 8 hours in culture. In this case, the alpha v and beta
3 integrin subunits were found at focal adhesions. After TPA treatment, HEL
cells deposited thrombospondin-immunoreactive material onto their culture
substratum, but synthesis of fibronectin, vitronectin, fibrinogen, or von
Willebrand factor was not detected. Thus, the results suggest that TPA
would activate several integrin receptors in HEL cells and also stimulate
the secretion of thrombospondin, which might be used as an adhesion ligand
for the integrin vitronectin receptor alpha v/beta 3 complex.
Volume 76,
Issue 3,
pp. 570-577,
08/01/1990
Copyright © 1990 by The American Society of Hematology
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