Characterization of the p130cas pathway in Ocular Cell Adhesions

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SV40-transformed human corneal epithelial cells (sCE), SV40-transformed human lens epithelial cells (sLE), normal human keratocyte (nK), human retinal pigment epithelial cell that stably expresses human telomerase reverse transcriptase (TERT-RPE) and human A-375 melanoma cells were used. Forth experiments were performed. First, A human corneal library was screened to isolate the human homologue of Crk-associated substrate(Cas). Second, after these cells attached on the fibronection (FN) or collagen I precoated culture plates, RGB peptides were added into culture medium and the morphological changes were observed. Third, the levels of tyrosine phosphorylation of FAK and Cas pre and after adding RGD peptides were evaluated in each cell. Forth, The expression of α2, α3, α5 andαv integrin subunit on each cell were analyzed by flow cytometryA clone, (H-2), encoding 3228 nucleotides, with a long open reading frame (870 amino acid residues) was isolated. We concluded that this clone was the human homologue of Cas. After addition of RGBS peptides, only human A-375 melanoma cells detached from the plate. RGD peptides induced tyrosine dephosphorylation of both FAK and Cas in A-375 melanoma cells but not in other ocular cells. While SCE significantly expressedα2 andα3 integrin and sLE, nK, and TERT-RPE doniinantly expressedα3 integrin, A-375 melanoma cells expressedα3 andα5 integrin. RGB peptides were effective in only target melanoma cell but not cells derived from human eye. This suggested that RGD peptides could be clinically applied to ocular melanoma