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Updated: Jul 15, 2026
In Vitro Assay to Measure Phosphatidylethanolamine Methyltransferase Activity
Published on: January 5, 2016
[Methyl-3H]-choline incorporation into MCF-7 cells: correlation with proliferation, choline kinase and phospholipase
Fatma Al-Saeedi1, Tim Smith, Andy Welch
1Department of Nuclear Medicine, Faculty of Medicine, Kuwait University, Safat, Kuwait. fatimas@hsc.edu.kw
Background:
[Methyl-3H]-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-3H]-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-3H]-PCho content expressed as pmol mg(-1) protein(-1) min(-1) (n = 6) was significantly higher in the exponentially growing (484.04 +/-2 0.23) 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.)/microg 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./microg protein (39.69 +/- 4.00%)) compared with confluent cells (99.10 +/- 1.35 d.p.m./microg 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.
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