[methyl-3H]Choline incorporation into MCF7 tumour cells: correlation with proliferation

F. Al-Saeedi, Andrew Welch, Timothy Andrew Davies Smith

Research output: Contribution to journalArticle

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Abstract

Purpose: The aim of the study was to investigate the intracellular location of [methyl-H-3]choline in MCF7 tumour cells and to determine the relationship between [methyl-H-3]choline incorporation and proliferation.

Methods: Tumour cells were incubated with [methyl-H-3]choline for 10 min, and then in cold medium to simulate the rapid blood clearance of [methyl-(11)pC]choline. Labelled metabolites were then extracted from cells by treating them with organic and aqueous solvents to determine the distribution of tracer between phospholipid and water-soluble metabolite pools. Aqueous extracts were subjected to thin-layer chromatography, ion exchange chromatography and a choline extraction procedure to identify H-3-containing metabolites. Procedures were carried out on fast- and slow-growing populations of MCF7 cells to determine the relationship between choline incorporation and proliferation.

Results: Only about 5% of [methyl-H-3]choline was present as phospholipid. [methyl-H-3]choline incorporation was found to be related to S-phase fraction. In another experiment, [methyl-C-14]choline incorporation was found to be correlated with [methyl-H-3]thymidine incorporation. The V-max of choline uptake was found to be increased whilst K-m was decreased in populations of MCF7 cells with higher proliferative fractions, compared with populations having lower proliferative fractions.

Conclusion: Choline incorporation into tumour cells under conditions that simulate rapid blood clearance of [methyl-C-11]choline is correlated with proliferation. Most of the activity (about 95%) was in the non-lipid fraction of the cell.

Original languageEnglish
Pages (from-to)660-667
Number of pages7
JournalEuropean Journal of Nuclear Medicine and Molecular Imaging
Volume32
Issue number6
DOIs
Publication statusPublished - Jun 2005

Keywords

  • choline
  • tumour
  • phosphocholine
  • PET
  • POSITRON-EMISSION-TOMOGRAPHY
  • ORGANIC CATION TRANSPORTERS
  • MITOGENIC GROWTH-FACTORS
  • CHOLINE KINASE-ACTIVITY
  • HUMAN BREAST-CANCER
  • PHOSPHOLIPID-METABOLISM
  • C-11 CHOLINE
  • BRAIN-TUMOR
  • P-31 NMR
  • PHOSPHATIDYLCHOLINE BIOSYNTHESIS

Cite this

[methyl-3H]Choline incorporation into MCF7 tumour cells: correlation with proliferation. / Al-Saeedi, F.; Welch, Andrew; Smith, Timothy Andrew Davies.

In: European Journal of Nuclear Medicine and Molecular Imaging, Vol. 32, No. 6, 06.2005, p. 660-667.

Research output: Contribution to journalArticle

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title = "[methyl-3H]Choline incorporation into MCF7 tumour cells: correlation with proliferation",
abstract = "Purpose: The aim of the study was to investigate the intracellular location of [methyl-H-3]choline in MCF7 tumour cells and to determine the relationship between [methyl-H-3]choline incorporation and proliferation.Methods: Tumour cells were incubated with [methyl-H-3]choline for 10 min, and then in cold medium to simulate the rapid blood clearance of [methyl-(11)pC]choline. Labelled metabolites were then extracted from cells by treating them with organic and aqueous solvents to determine the distribution of tracer between phospholipid and water-soluble metabolite pools. Aqueous extracts were subjected to thin-layer chromatography, ion exchange chromatography and a choline extraction procedure to identify H-3-containing metabolites. Procedures were carried out on fast- and slow-growing populations of MCF7 cells to determine the relationship between choline incorporation and proliferation.Results: Only about 5{\%} of [methyl-H-3]choline was present as phospholipid. [methyl-H-3]choline incorporation was found to be related to S-phase fraction. In another experiment, [methyl-C-14]choline incorporation was found to be correlated with [methyl-H-3]thymidine incorporation. The V-max of choline uptake was found to be increased whilst K-m was decreased in populations of MCF7 cells with higher proliferative fractions, compared with populations having lower proliferative fractions.Conclusion: Choline incorporation into tumour cells under conditions that simulate rapid blood clearance of [methyl-C-11]choline is correlated with proliferation. Most of the activity (about 95{\%}) was in the non-lipid fraction of the cell.",
keywords = "choline, tumour, phosphocholine, PET, POSITRON-EMISSION-TOMOGRAPHY, ORGANIC CATION TRANSPORTERS, MITOGENIC GROWTH-FACTORS, CHOLINE KINASE-ACTIVITY, HUMAN BREAST-CANCER, PHOSPHOLIPID-METABOLISM, C-11 CHOLINE, BRAIN-TUMOR, P-31 NMR, PHOSPHATIDYLCHOLINE BIOSYNTHESIS",
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T1 - [methyl-3H]Choline incorporation into MCF7 tumour cells: correlation with proliferation

AU - Al-Saeedi, F.

AU - Welch, Andrew

AU - Smith, Timothy Andrew Davies

PY - 2005/6

Y1 - 2005/6

N2 - Purpose: The aim of the study was to investigate the intracellular location of [methyl-H-3]choline in MCF7 tumour cells and to determine the relationship between [methyl-H-3]choline incorporation and proliferation.Methods: Tumour cells were incubated with [methyl-H-3]choline for 10 min, and then in cold medium to simulate the rapid blood clearance of [methyl-(11)pC]choline. Labelled metabolites were then extracted from cells by treating them with organic and aqueous solvents to determine the distribution of tracer between phospholipid and water-soluble metabolite pools. Aqueous extracts were subjected to thin-layer chromatography, ion exchange chromatography and a choline extraction procedure to identify H-3-containing metabolites. Procedures were carried out on fast- and slow-growing populations of MCF7 cells to determine the relationship between choline incorporation and proliferation.Results: Only about 5% of [methyl-H-3]choline was present as phospholipid. [methyl-H-3]choline incorporation was found to be related to S-phase fraction. In another experiment, [methyl-C-14]choline incorporation was found to be correlated with [methyl-H-3]thymidine incorporation. The V-max of choline uptake was found to be increased whilst K-m was decreased in populations of MCF7 cells with higher proliferative fractions, compared with populations having lower proliferative fractions.Conclusion: Choline incorporation into tumour cells under conditions that simulate rapid blood clearance of [methyl-C-11]choline is correlated with proliferation. Most of the activity (about 95%) was in the non-lipid fraction of the cell.

AB - Purpose: The aim of the study was to investigate the intracellular location of [methyl-H-3]choline in MCF7 tumour cells and to determine the relationship between [methyl-H-3]choline incorporation and proliferation.Methods: Tumour cells were incubated with [methyl-H-3]choline for 10 min, and then in cold medium to simulate the rapid blood clearance of [methyl-(11)pC]choline. Labelled metabolites were then extracted from cells by treating them with organic and aqueous solvents to determine the distribution of tracer between phospholipid and water-soluble metabolite pools. Aqueous extracts were subjected to thin-layer chromatography, ion exchange chromatography and a choline extraction procedure to identify H-3-containing metabolites. Procedures were carried out on fast- and slow-growing populations of MCF7 cells to determine the relationship between choline incorporation and proliferation.Results: Only about 5% of [methyl-H-3]choline was present as phospholipid. [methyl-H-3]choline incorporation was found to be related to S-phase fraction. In another experiment, [methyl-C-14]choline incorporation was found to be correlated with [methyl-H-3]thymidine incorporation. The V-max of choline uptake was found to be increased whilst K-m was decreased in populations of MCF7 cells with higher proliferative fractions, compared with populations having lower proliferative fractions.Conclusion: Choline incorporation into tumour cells under conditions that simulate rapid blood clearance of [methyl-C-11]choline is correlated with proliferation. Most of the activity (about 95%) was in the non-lipid fraction of the cell.

KW - choline

KW - tumour

KW - phosphocholine

KW - PET

KW - POSITRON-EMISSION-TOMOGRAPHY

KW - ORGANIC CATION TRANSPORTERS

KW - MITOGENIC GROWTH-FACTORS

KW - CHOLINE KINASE-ACTIVITY

KW - HUMAN BREAST-CANCER

KW - PHOSPHOLIPID-METABOLISM

KW - C-11 CHOLINE

KW - BRAIN-TUMOR

KW - P-31 NMR

KW - PHOSPHATIDYLCHOLINE BIOSYNTHESIS

U2 - 10.1007/S00259-004-1707-6

DO - 10.1007/S00259-004-1707-6

M3 - Article

VL - 32

SP - 660

EP - 667

JO - European Journal of Nuclear Medicine and Molecular Imaging

JF - European Journal of Nuclear Medicine and Molecular Imaging

SN - 1619-7070

IS - 6

ER -