Changes in glycosaminoglycan structure and composition of the main heparan sulphate proteoglycan from human colon carcinoma cells (perlecan) during cell differentiation

Abstract

Colon carcinoma cells provide a useful model to study the biochemical processes associated with cell differentiation. Undifferentiated HT29, differentiated HT29MTXϪ3 and HT29MTX Ϫ6 , and Caco2 human colon carcinoma cells have been used to study the production of proteoglycans and to characterize the glycosaminoglycan structure of the heparan sulphate chains. All the cell lines produce mainly a heparan sulphate proteoglycan that is found partly in the extracellular medium and associated to the cell membrane. The heparan sulphate proteoglycans from the media were purified by ion-exchange chromatography and subjected to structural analysis. The heparan sulphate proteoglycan from differentiated cells is larger and more homogeneous in size than the heparan sulphate proteoglycan from undifferentiated HT29 cells. No differences in protein core structure were observed when cells were labeled with [35 S]methionine and the protein cores visualized by gel electrophoresis. Nevertheless, differences in glycosaminoglycan composition were found correlated with the degree of differentiation. The heparan sulphate chains from differentiated HT29MTXϪ3 and HT29MTX Ϫ6 cells have a higher sulphation degree than those from undifferentiated HT29 cells. The heparan sulphate from Caco2 cells is the most highly sulphated species. The differences are mainly attributed to O-sulphate groups. The increase in O-sulphation was more pronounced for D-glucosamine 6-O-sulphate than for L-iduronic acid 2-O-sulphate groups.Keywords : proteoglycan ; glycosaminoglycan; colon carcinoma; perlecan; cell differentiation.Sulphated glycosaminoglycans, and particularly heparan sul-changes in protein glycosylation occur during the process of cell differentiation in the HT29 cell line (Wice et al., 1985; Ogierphate (HS), play an important role in cellϪcell and cellϪsub-strate interactions, cell attachment and spreading, growth factor Denis et al., 1988). Among the other established colon carcinoma cell lines, the Caco2 cell line has been widely used beregulation and cell transformation (Gallagher, 1989; Kjellén and Lindahl, 1991;Hardingham and Fosang, 1992; Yanagishita and cause it spontaneously expresses a very high degree of enterocytic differentiation (Rousset, 1986). Hascall, 1992). In particular, alterations in proteoglycan composition and/or structure have been described in several transHeparan sulphate proteoglycans have been implicated in a number of fundamental cell-biological phenomena. The HS side formed cells (Robinson et al., 1984;Keller et al., 1980; Lévy et al., 1990;Knudson et al., 1990;Pejler and David, 1987; Winter-chains have the ability to bind, in a more or less specific fashion, to a variety of proteins, including extracellular matrix proteins, bourne and Mora, 1981). A good model to investigate the relationship between proteoglycan structure and cell differentiation enzymes, enzyme inhibitors, growth factors and other cytokines (Gallagher, 1989;Spillmann and Lindahl, 1994; Salmivirta et would be a cell line with a wide range of phenotyp...

Distribution and Clinical Significance of Heparan Sulfate Proteoglycans in Ovarian Cancer

“…Thus, protein expression and turnover may remain stable but HSPG function may be altered by structural changes in the heparan sulfate chains. This has been observed in an experimental model of colon cancer, where transition from adenoma to carcinoma is accompanied by altered heparan sulfate structure that in turn changes reactivity to fibroblast growth factors (41)(42)(43). Similarly, heparan sulfate is subject to degradation by proteolytic enzymes called heparanases, which contribute to tumor cell invasion and metastasis by liberating fragments of heparan sulfate that can activate growth factors (20).…”

Mucin Granule Intraluminal Organization in Living Mucous/Goblet Cells

“…3, E and F) might contribute to limit protein intragranular diffusion. Nevertheless, although heparan sulfate is known to be expressed in HT29 cells (50), the existence of intragranular proteoglycans in mucous/goblet cells remains unproven. Contrary to inhibition of sulfation, reduced sialylation diminished SHGFP-MUC5AC/CK intragranular mobility.…”

Requirement of both mucins and proteoglycans in cell‐cell dissociation and invasiveness of colon carcinoma HT‐29 cells

“…Matrix and cell surface proteoglycans carrying heparan sulfate glycosaminoglycans have been characterized in HT-29 cells. [33][34][35] We thus investigated the effect of an antibody directed against the heparan sulfate moiety. This antibody displayed a similar effect as the anti-MUC1 and anti-MUC5AC antibody ( Fig.…”

“…Perlecan was described as a main proteoheparan sulfate of HT-29 cells, and a variability in its carbohydrate structure was shown according to the differentiation state of HT-29 cells. 34,35 In this regard, invasive cells of mouse or dog epithelial cell lines were previously described to contain enlarged proteoglycans in comparison to non invasive cells, and these enlarged proteoglycans were proposed to disturb the function of E-cadherin through steric hindrance. 17 However, this work, which for the first time combines a study of mucins and proteoglycans, shows that the role of HSPGs in the invasive behavior of HT-29 5M21 cells is shared with MUC1 and MUC5AC.…”