Differential expression of sialic acid and N-acetylgalactosamine residues on the cell surface of intestinal epithelial cells according to normal or metastatic potential.
In this study we investigated the levels of expression of sialic acid and N-acetylgalactosamine residues on the cell surface of a normal intestinal epithelium cell line, IEC-6, and in two colon adenocarcinoma cell lines with different metastatic potential, Caco-2 and HCT-116. Glycoprotein expression was estimated initially by cytochemistry with WGA and HPA lectins and biochemistry with isolated plasma membrane fractions of the cells. Fluorescence and electron microscopic analyses revealed differences in the expression profile of carbohydrates recognized by the lectins used on the cell surface of IEC-6, Caco-2, and HCT-116 cells. Lectin blotting identified a range of eight HPA-binding glycoprotein bands with molecular weights of 16-66 kD in Caco-2 cells, six glycoproteins of 16-36 kD, and three protein bands of 35, 24, and 21 kD in IEC-6 cells. A minor band of 66 kD and a major one of 50 kD for WGA-binding glycoproteins were observed in IEC-6 cells and seven glycoproteins of 18-97 kD in Caco-2 and HCT-116 cells but with a visible higher expression of these glycoproteins in the latter. Furthermore, significant quantitative difference in levels of expression of WGA- but not of HPA-binding glycoconjugates was noted, as analyzed by high-resolution scanning electron microscopy using backscattered electron images of cells incubated with gold-labeled lectins.
de Albuquerque Garcia Redondo
Patricia
P
Divisão de Biologia Celular, Grupo de Biologia Estrutural, Centro de Pesquisa, Instituto Nacional de Câncer, Rio de Janeiro, Brasil.
Nakamura
Celso Vataru
CV
de Souza
Wanderley
W
Morgado-Díaz
José Andrés
JA
D000116:Acetylgalactosamine / Q000378:metabolism*; D000230:Adenocarcinoma / Q000378:metabolism* / Q000473:pathology; D000818:Animals; D018938:Caco-2 Cells; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000378:metabolism* / Q000473:pathology; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007202:Indicators and Reagents; D007413:Intestinal Mucosa / Q000166:cytology* / Q000378:metabolism* / Q000473:pathology; D037102:Lectins; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D019158:N-Acetylneuraminic Acid / Q000378:metabolism*; D009362:Neoplasm Metastasis; D051381:Rats
D007202:Indicators and Reagents; D037102:Lectins; D019158:N-Acetylneuraminic Acid; D000116:Acetylgalactosamine