Alteration of lacto-series glycolipid glycosyltransferase activities in human colonic adenocarcinoma DLD-1 cells after culture in N,N-dimethylformamide-containing medium.

Abstract

Human colonic adenocarcinoma DLD-1 cells were grown under conditions which induce characteristics of differentiated cells using medium containing 0.8% N,N-dimethylformamide in order to study alterations in glycosphingolipid glycosyltransferase activities during this process. Analysis of biosynthetic reactions involved in lacto-series antigen synthesis revealed no changes in the specific activities of either beta 1----4galactosyltransferase or alpha 1----3/4fucosyltransferase with N,N-dimethylformamide treatment. However, a dramatic decrease of from 14- to 20-fold in the beta 1----3N-acetylglucosaminyltransferase activity was observed in the treated cells. This enzyme catalyzes the rate-limiting step in lacto-series core chain synthesis. This is consistent with the pattern of regulation of lacto-series antigen expression found to occur during oncogenesis in human colonic mucosa (Holmes EH, Hakomori S, Ostrander GK: J Biol Chem 262:15649, 1987). Total glycolipids from untreated and N,N-dimethylformamide-treated cells were isolated and subjected to TLC immunostain analysis and solid phase radioimmunoassay with a series of monoclonal antibodies specific for lacto-series-based carbohydrate antigens. A decrease of about 2-fold or less in the quantity of lacto-series antigens was observed as a consequence of N,N-dimethylformamide treatment in both neutral glycolipid and ganglioside fractions. The results suggest that only very low levels of beta 1----3N-acetylglucosaminyltransferase activity are required for the steady state expression of significant levels of lacto-series based glycolipids and that modulation of its activity levels by N,N-dimethylformamide treatment in DLD-1 cells represents a convenient in vitro system for studying aspects of regulation of lacto-series antigen expression.

• Authors: E H Holmes, T G Greene
• Journal: Journal of cellular biochemistry (J Cell Biochem) Vol. 44 Issue 2 Pg. 93-105 (Oct 1990) ISSN: 0730-2312 [Print] United States
• PMID: 2250046 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Antigens, Neoplasm
• Glycolipids
• Neoplasm Proteins
• Dimethylformamide
• Hexosyltransferases

Topics

• Adenocarcinoma (pathology)
• Antigens, Neoplasm (biosynthesis)
• Cell Differentiation (drug effects)
• Cell Transformation, Neoplastic
• Colonic Neoplasms (pathology)
• Dimethylformamide (pharmacology)
• Glycolipids (metabolism)
• Glycosylation
• Hexosyltransferases (analysis)
• Humans
• Neoplasm Proteins (analysis)
• Tumor Cells, Cultured (drug effects, enzymology, pathology)