Metabolic changes in undifferentiated and differentiated human colon adenocarcinoma cells studied by multinuclear magnetic resonance spectroscopy

Jean-Philippe Galons, Jacques Fantini, Jean Vion-Dury, Patrick J. Cozzone, Paul Canioni

Research output: Contribution to journalArticle

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Abstract

Aspects of energetic and intermediary metabolism were studied in a colon adenocarcinoma cell line (HT29) by multinuclear magnetic resonance spectroscopy. Experiments were carried out on the HT29-D4 clone, which was isolated by limit dilution techniques. This clone, usually undifferentiated (D4-UD), can be maintained in a differentiated state (D4-D) in a glucose-free medium. Metabolic data were obtained by NMR analysis of perchloric acid extracts from D4-UD and D4-D cells. Phosphorus-31 and proton NMR spectra showed the presence of a large amount of choline and phosphorylcholine in the differentiated state (400% and 200%, respectively, of the levels found in D4-UD cells). Other differences appeared in the content of phosphocreatine (absent in D4-D cells) and myoinositol (absent in D4-UD cells). Carbon-13 spectra were recorded from perchloric acid extracts of cells incubated with [1-13C]-labeled glucose or [2-13C]-labeled acetate. The data indicated that both types of cells metabolize glucose through the glycolytic pathway to give lactate, but only D4-D cells were able to store glucose as glycogen at a very high level. A mathematical analysis of fluxes through the tricarboxylic acid (TCA) cycle was developed on the basis of models derived from previous 14C tracer studies. The model was based on the steady-state labeling of glutamate carbons by the 13C isotope and gave the fraction of labeled acetyl-Coa entering the TCA cycle, and the activity of anaplerotic reactions relative to the flux through the citrate synthetase reaction. The data indicated that y > 0.3 in all cases. Only 15% and 30% of labeled acetyl CoA entered the TCA cycle in D4-UD and D4-D cells, respectively, under labeled glucose incubation: these values were significantly different upon labeled acetate feeding, reaching 55% for D4-UD cells and 85% for D4-D cells. The main result of this study is that the process of differentiation of HT29 cells is correlated with a large increase in the activity of oxidative metabolism.

Original languageEnglish (US)
Pages (from-to)949-961
Number of pages13
JournalBiochimie
Volume71
Issue number8
DOIs
StatePublished - 1989
Externally publishedYes

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Magnetic resonance spectroscopy
Somatostatin-Secreting Cells
Colon
Adenocarcinoma
Magnetic Resonance Spectroscopy
Citric Acid Cycle
Glucose
Metabolism
Acetates
Carbon
Clone Cells
Nuclear magnetic resonance
Carbon Isotopes
Fluxes
Indicator Dilution Techniques
HT29 Cells
Acetyl Coenzyme A
Phosphorylcholine
Phosphocreatine
Inositol

Keywords

  • differentiation
  • HT29
  • metabolism
  • NMR-cultured cells

ASJC Scopus subject areas

  • Biochemistry

Cite this

Metabolic changes in undifferentiated and differentiated human colon adenocarcinoma cells studied by multinuclear magnetic resonance spectroscopy. / Galons, Jean-Philippe; Fantini, Jacques; Vion-Dury, Jean; Cozzone, Patrick J.; Canioni, Paul.

In: Biochimie, Vol. 71, No. 8, 1989, p. 949-961.

Research output: Contribution to journalArticle

Galons, Jean-Philippe ; Fantini, Jacques ; Vion-Dury, Jean ; Cozzone, Patrick J. ; Canioni, Paul. / Metabolic changes in undifferentiated and differentiated human colon adenocarcinoma cells studied by multinuclear magnetic resonance spectroscopy. In: Biochimie. 1989 ; Vol. 71, No. 8. pp. 949-961.
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