Related Experiment Videos
Changes in glycosylation of L1210 cells after exposure to various antimetabolites
T W De Graaf1, S S Slot, G J Peters
1Department of Medical Chemistry, Faculty of Medicine, Vrije Universiteit, Amsterdam, The Netherlands.
Abstract:
This study establishes that antimetabolites do have the potency to change cellular glycosylation, as was suggested in our previous review (Eur J Cancer 1990, 26, 516-523). Murine leukaemia L1210 cells were exposed to various antimetabolites under non-lethal conditions. The antimetabolites 5-fluorouracil (5FU), arabinofuranosylcytosine (AraC), methotrexate (MTX) and 6-mercaptopurine (6MP), but not 6-thioguanine, induced considerable changes in the metabolic incorporation of radioactively labelled monosaccharides. Each antimetabolite exhibited a different effect. Significant differences were found between the radioactivity incorporated from the monosaccharides glucosamine, fucose, mannose and galactose, relative to control values. Polyacrylamide gel electrophoresis indicated that changes were induced in the glycosylation of individual glycoproteins. 5FU, AraC, MTX and 6MP all influenced both pyrimidine- and purine-mediated sugar incorporation. This excludes, therefore, direct effects of the antimetabolites on their analogue nucleotide-sugars. The antimetabolite-induced changes in glycosylation did not directly correlate with the observed cell-cycle effects of the antimetabolites.
Insights
Antimetabolites like 5-fluorouracil (5FU) can alter cellular glycosylation by affecting sugar incorporation into glycoproteins. These changes in cancer cells do not directly correlate with cell-cycle effects.
Area of Science:
- Biochemistry
- Cell Biology
- Cancer Research
Background:
- Cellular glycosylation is a critical process involved in various biological functions.
- Antimetabolites are a class of drugs used in cancer chemotherapy.
- Previous research suggested a potential link between antimetabolites and cellular glycosylation.
Purpose of the Study:
- To investigate the effects of specific antimetabolites on cellular glycosylation in murine leukemia L1210 cells.
- To determine if antimetabolites alter the metabolic incorporation of monosaccharides.
- To examine the impact on glycoprotein glycosylation patterns.
Main Methods:
- Exposure of L1210 cells to non-lethal concentrations of antimetabolites: 5-fluorouracil (5FU), arabinofuranosylcytosine (AraC), methotrexate (MTX), 6-mercaptopurine (6MP), and 6-thioguanine.
- Measurement of metabolic incorporation of radioactively labeled monosaccharides (glucosamine, fucose, mannose, galactose).
- Analysis of glycoprotein changes using polyacrylamide gel electrophoresis.
Main Results:
- 5FU, AraC, MTX, and 6MP significantly altered the incorporation of labeled monosaccharides, indicating changes in cellular glycosylation.
- Each antimetabolite demonstrated a distinct effect on sugar incorporation.
- Changes in glycoprotein glycosylation were observed.
- The observed glycosylation changes were independent of direct effects on nucleotide-sugar analogues and did not correlate with cell-cycle effects.
Conclusions:
- Antimetabolites possess the ability to modify cellular glycosylation patterns.
- The mechanism involves influencing both pyrimidine- and purine-mediated sugar incorporation.
- Glycosylation alterations induced by these antimetabolites are distinct from their cell-cycle effects.