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Abstract
PURPOSE: 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.