The decreased adhesion of Y79 retinoblastoma cells to extracellular matrix proteins is due to a deficit of integrin receptors.
OBJECTIVE
This study was designed to determine whether the Y79 retinoblastoma cell line, a prototype for retinoblastoma cells, exhibits differential adhesive properties toward extracellular matrix (ECM)/basement membrane (BM) proteins compared to normal human retinal (NHR) cells. A second goal was to determine whether differences in adhesion are related to differences in the expression of integrin subunits.


METHODS
Y79 cells and NHR cells were tested for their ability to adhere and spread in microtiter wells adsorbed with the ECM/BM proteins laminin, fibronectin, and type IV collagen, as well as fragments of these proteins. The presence of cell surface integrins was determined by flow cytometry, using monoclonal antibodies (mAbs) against integrin subunits. For inhibition assays, cells were preincubated with mAbs against integrin subunits before the adhesion assays.


RESULTS
NHR cells adhered to and spread on laminin, fibronectin, and type IV collagen, whereas Y79 cells only adhered moderately to fibronectin. NHR cells expressed high levels of beta 1, alpha 1, alpha 2, alpha 3, alpha 4, alpha 5, alpha 6, and alpha v integrin subunits, and they used these integrin subunits to adhere to all three ECM proteins. In contrast, Y79 cells expressed high levels of only the alpha 4 and beta 1 integrin subunits, used to adhere to fibronectin.


CONCLUSIONS
Y79 cells have decreased adhesive capabilities toward ECM/BM proteins, compared to NHR cells. These differences can be attributed, in part, to their significantly lower levels of alpha 1, alpha 2, alpha 3, and alpha 5 integrin subunits, which serve as receptors for type IV collagen, laminin, and fibronectin.
Skubitz
A P
AP
Department of Laboratory Medicine, University of Minnesota Medical School, Minneapolis 55455.
Grossman
M D
MD
McCarthy
J B
JB
Wayner
E A
EA
Cameron
J D
JD
D000911:Antibodies, Monoclonal; D001485:Basement Membrane / Q000378:metabolism; D002448:Cell Adhesion; D002455:Cell Division; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D005134:Eye Neoplasms / Q000378:metabolism* / Q000473:pathology; D005434:Flow Cytometry; D006801:Humans; D016023:Integrins / Q000172:deficiency / Q000378:metabolism*; D016024:Receptors, Cytoadhesin / Q000378:metabolism*; D012160:Retina / Q000166:cytology / Q000378:metabolism; D012175:Retinoblastoma / Q000378:metabolism* / Q000473:pathology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D016326:Extracellular Matrix Proteins; D016023:Integrins; D016024:Receptors, Cytoadhesin