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Differential Expression of Sialic Acid and N-acetylgalactosamine Residues on the Cell Surface of Intestinal Epithelial Cells According to Normal or Metastatic Potential

Patricia de Albuquerque Garcia Redondo, Celso Vataru Nakamura, Wanderley de Souza and José Andrés Morgado–Díaz

Divisão de Biologia Celular, Grupo de Biologia Estrutural, Centro de Pesquisa, Instituto Nacional de Câncer, Rio de Janeiro, Brasil (PdAGR,JAM–D); Laboratório de Microbiologia, Departamento de Análises Clínicas, Universidade Estadual de Maringá, PR, Brasil (CVN); and Laboratório de Ultraestrutura Celular Hertha Meyer, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brasil (WdS)

Correspondence to: José Andrés Morgado–Díaz, Divisão de Biologia Celular, Grupo de Biologia Estrutural, Centro de Pesquisa, Instituto Nacional de Câncer, Rua André Cavalcanti 37, 5° Andar, 20231-050, Rio de Janeiro, Brasil. E-mail: jmorgado{at}inca.gov.br

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. (J Histochem Cytochem 52:629–640, 2004)

Key Words: lectin cytochemistry • sialic acid • N-acetylgalactosamine • epithelial cells • colon • adenocarcinoma

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