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Synthesis of Glycosaminoglycans by Undifferentiated and Differentiated HT29 Human Colonic Cancer Cells¹

INSERM Unitè 61, Biologie Cellulaire et Physiopathologie Digestives, 3 avenue Molière, 67200 Strasbourg Hautepierre, France

Among the extracellular matrix components which have been suggested to be involved in developmental and neoplastic changes are glycosaminoglycans (GAGs). To try to correlate their amount and nature with the process of enterocytic differentiation, we studied glycosaminoglycan synthesis of human colonic adenocarcinoma cells (HT29 cell line) by [³H]glucosamine and [³⁵S]sulfate incorporation. Enterocytic differentiation of the cells obtained in a sugar-free medium (for review, see A. Zweibaum et al. In: Handbook of Physiology. Intestinal Transport of the Gastrointestinal System, in press, 1987) resulted in a marked increase in total incorporation of labeled precursors (20-fold for [³H]glucosamine, 4.5-fold for [³⁵S]sulfate) as well as in uronic acid content (5-fold); most of the synthesized GAGs were found associated with the cell pellet. Chromatographic and electrophoretic analysis of the labeled GAGs revealed that undifferentiated cells synthesized and secreted hyaluronic acid, heparan sulfate, and one class of chondroitin sulfate. Differentiation of HT29 cells became associated with the synthesis of an additional class of chondroitin sulfate (CS4) concomitant to a decrease in heparan sulfate which is no longer found secreted in the medium. Furthermore the charge density of this latter GAG component varied as assessed by a shift of its affinity on ion-exchange chromatography.

¹ Financial support was given by INSERM, ARC (Grant 6236), and The Ligue Nationale Française contre le Cancer.

² To whom requests for reprints should be addressed.

³ Recipient of a fellowship from ARC.

Received 11/ 5/86. Revised 3/30/87. Accepted 5/19/87.

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