Adhesive properties and integrin expression profiles of two colonic cancer populations differing by their spreading on laminin

Abstract

The mostly undifferentiated parental HT29 (HT29p) human colonic adenocarcinoma cell line and a differentiated subpopulation selected by the anti-cancer drug 5-fluorouracil (HT29-Fu) (Lesuffleur et al. (1991) Int. J. Cancer 49, 721-730) display strikingly different behavior when grown on laminin coatings: the former grows as aggregates while the latter grows as monolayers. In an attempt to explain this difference, we performed a comparative study of cell adhesion properties and of expression, involvement and localization of the alpha 6, beta 1 and beta 4 subunits constituting the integrin family among the two cell populations. HT29p and HT29-Fu cells exhibited a similar adhesion pattern to laminin and laminin fragments E8 and P1. In both cell lines, cell adhesion could be blocked at about 90% with anti-alpha 6 subunit antibodies and around 30-50% with anti-beta 1 antibodies; no inhibition of the cell adhesion was obvious when using anti-beta 4 antibodies. Immunoprecipitations of iodinated membrane-solubilized proteins and immunoblotting experiments showed that all alpha 6 chains expressed in both HT29p and HT29-Fu cell populations exist as alpha 6 beta 4 integrins; beta 1 subunits are associated with alpha 2 and alpha 3 chains. When HT29p or HT29-Fu cells were injected subcutaneously in nude mice, a similar expression pattern of alpha 6, beta 4 and beta 1 integrin subunits was noticeable in the resulting tumors: alpha 6 and beta 4 subunits were localized at the basal surface of the tumor cells facing the stromal elements, and to a lesser extent at the cell-cell contacts within the tumor-cell clumps; beta 1 subunits were mainly found within the cytoplasm of the tumor cells. Despite these overall similarities among the two cell lines, the following changes could account for their different behavior on laminin: less proteolytic processing of the beta 4 integrin subunit occurred in HT29-Fu cells yielding peptidic fragments of 175 kDa, which are absent from the parental cells; the immunostaining pattern of the various subunits demonstrated a segregation of alpha 6, beta 4 and beta 1 integrin subunits on the basal side of the HT29-Fu cells when cultured on laminin to the detriment of their lateral location, a phenomenon that was not obvious in the parental cells. Altogether, these results suggest that the distinct behavior of the undifferentiated versus differentiated HT29 cell populations on laminin is not related to altered adhesion properties of the cells but rather to a deficient stabilization of the adhesion leading to cell spreading.(ABSTRACT TRUNCATED AT 400 WORDS)