[Methyl-3H]-choline incorporation into MCF-7 Cells: Correlation with Proliferation-Choline Kinase and Phospholipase D Assay

Fatma Al-Saeedi, Tim Smith, Andy Welch

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Background: [Methyl-H-3]-choline is a promising new positron emission tomography (PET) agent used for cancer imaging whose mechanism has still not fully been elucidated. In this study, whether [methyl-H-3]-choline determined by measuring the activity of choline kinase (ChoK) and phospholipase D (PLD) in rapidly proliferating and confluent breast cancer MCF-7 cells is related to cell proliferation or not was investigated. Materials and Methods: The activity of ChoK and PLD were determined using ion exchange chromatography and transphosphatidylation assay respectively. Results: [Methyl-H-3]-PCho content expressed as pmol mg(-1) protein(-1) min(-1) (n=6) was significantly higher in the exponentially growing (484.04 +/- 20.23) compared with confluent (70.35 +/- 9.83) cells using Student's t-test (p < 0.001). Moreover, PLD activity expressed as the mean (n=6) (disintegration per minute (d.p.m.)/mu g protein +/- SD (mean S phase SD)) showed significantly higher (p < 0.001) activity in the exponentially growing cells (196.39 +/- 2.21 d.p.m./mu g protein (39.69 +/- 4. 00%)) compared with confluent cells (99.10 +/- 1.35 d.p.m./mu g protein (9.33 +/- 0.82%)). Conclusion: This study indicates that the major water-soluble choline metabolite was phosphocholine (PCho) as a consequence of increased ChoK and PLD activity in the exponentially growing cells compared to confluent cells.

Original languageEnglish
Pages (from-to)901-906
Number of pages6
JournalAnticancer Research
Volume27
Issue number2
Publication statusPublished - 2007

Keywords

  • radiopharmaceuticals
  • choline
  • phosphocholine
  • PLD
  • PET
  • phospholipase D
  • mitogenic growth-factors
  • breast-cancer
  • phosphatidylcholine breakdown
  • signal-transduction
  • 2ND messenger
  • RAS oncogene
  • metabolism
  • diacylglycerol
  • overexpression

Cite this

[Methyl-3H]-choline incorporation into MCF-7 Cells : Correlation with Proliferation-Choline Kinase and Phospholipase D Assay. / Al-Saeedi, Fatma; Smith, Tim; Welch, Andy.

In: Anticancer Research, Vol. 27, No. 2, 2007, p. 901-906.

Research output: Contribution to journalArticle

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title = "[Methyl-3H]-choline incorporation into MCF-7 Cells: Correlation with Proliferation-Choline Kinase and Phospholipase D Assay",
abstract = "Background: [Methyl-H-3]-choline is a promising new positron emission tomography (PET) agent used for cancer imaging whose mechanism has still not fully been elucidated. In this study, whether [methyl-H-3]-choline determined by measuring the activity of choline kinase (ChoK) and phospholipase D (PLD) in rapidly proliferating and confluent breast cancer MCF-7 cells is related to cell proliferation or not was investigated. Materials and Methods: The activity of ChoK and PLD were determined using ion exchange chromatography and transphosphatidylation assay respectively. Results: [Methyl-H-3]-PCho content expressed as pmol mg(-1) protein(-1) min(-1) (n=6) was significantly higher in the exponentially growing (484.04 +/- 20.23) compared with confluent (70.35 +/- 9.83) cells using Student's t-test (p < 0.001). Moreover, PLD activity expressed as the mean (n=6) (disintegration per minute (d.p.m.)/mu g protein +/- SD (mean S phase SD)) showed significantly higher (p < 0.001) activity in the exponentially growing cells (196.39 +/- 2.21 d.p.m./mu g protein (39.69 +/- 4. 00{\%})) compared with confluent cells (99.10 +/- 1.35 d.p.m./mu g protein (9.33 +/- 0.82{\%})). Conclusion: This study indicates that the major water-soluble choline metabolite was phosphocholine (PCho) as a consequence of increased ChoK and PLD activity in the exponentially growing cells compared to confluent cells.",
keywords = "radiopharmaceuticals, choline, phosphocholine, PLD, PET, phospholipase D, mitogenic growth-factors, breast-cancer, phosphatidylcholine breakdown, signal-transduction, 2ND messenger, RAS oncogene, metabolism, diacylglycerol, overexpression",
author = "Fatma Al-Saeedi and Tim Smith and Andy Welch",
year = "2007",
language = "English",
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pages = "901--906",
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TY - JOUR

T1 - [Methyl-3H]-choline incorporation into MCF-7 Cells

T2 - Correlation with Proliferation-Choline Kinase and Phospholipase D Assay

AU - Al-Saeedi, Fatma

AU - Smith, Tim

AU - Welch, Andy

PY - 2007

Y1 - 2007

N2 - Background: [Methyl-H-3]-choline is a promising new positron emission tomography (PET) agent used for cancer imaging whose mechanism has still not fully been elucidated. In this study, whether [methyl-H-3]-choline determined by measuring the activity of choline kinase (ChoK) and phospholipase D (PLD) in rapidly proliferating and confluent breast cancer MCF-7 cells is related to cell proliferation or not was investigated. Materials and Methods: The activity of ChoK and PLD were determined using ion exchange chromatography and transphosphatidylation assay respectively. Results: [Methyl-H-3]-PCho content expressed as pmol mg(-1) protein(-1) min(-1) (n=6) was significantly higher in the exponentially growing (484.04 +/- 20.23) compared with confluent (70.35 +/- 9.83) cells using Student's t-test (p < 0.001). Moreover, PLD activity expressed as the mean (n=6) (disintegration per minute (d.p.m.)/mu g protein +/- SD (mean S phase SD)) showed significantly higher (p < 0.001) activity in the exponentially growing cells (196.39 +/- 2.21 d.p.m./mu g protein (39.69 +/- 4. 00%)) compared with confluent cells (99.10 +/- 1.35 d.p.m./mu g protein (9.33 +/- 0.82%)). Conclusion: This study indicates that the major water-soluble choline metabolite was phosphocholine (PCho) as a consequence of increased ChoK and PLD activity in the exponentially growing cells compared to confluent cells.

AB - Background: [Methyl-H-3]-choline is a promising new positron emission tomography (PET) agent used for cancer imaging whose mechanism has still not fully been elucidated. In this study, whether [methyl-H-3]-choline determined by measuring the activity of choline kinase (ChoK) and phospholipase D (PLD) in rapidly proliferating and confluent breast cancer MCF-7 cells is related to cell proliferation or not was investigated. Materials and Methods: The activity of ChoK and PLD were determined using ion exchange chromatography and transphosphatidylation assay respectively. Results: [Methyl-H-3]-PCho content expressed as pmol mg(-1) protein(-1) min(-1) (n=6) was significantly higher in the exponentially growing (484.04 +/- 20.23) compared with confluent (70.35 +/- 9.83) cells using Student's t-test (p < 0.001). Moreover, PLD activity expressed as the mean (n=6) (disintegration per minute (d.p.m.)/mu g protein +/- SD (mean S phase SD)) showed significantly higher (p < 0.001) activity in the exponentially growing cells (196.39 +/- 2.21 d.p.m./mu g protein (39.69 +/- 4. 00%)) compared with confluent cells (99.10 +/- 1.35 d.p.m./mu g protein (9.33 +/- 0.82%)). Conclusion: This study indicates that the major water-soluble choline metabolite was phosphocholine (PCho) as a consequence of increased ChoK and PLD activity in the exponentially growing cells compared to confluent cells.

KW - radiopharmaceuticals

KW - choline

KW - phosphocholine

KW - PLD

KW - PET

KW - phospholipase D

KW - mitogenic growth-factors

KW - breast-cancer

KW - phosphatidylcholine breakdown

KW - signal-transduction

KW - 2ND messenger

KW - RAS oncogene

KW - metabolism

KW - diacylglycerol

KW - overexpression

M3 - Article

VL - 27

SP - 901

EP - 906

JO - Anticancer Research

JF - Anticancer Research

SN - 0250-7005

IS - 2

ER -