Biodisposition and metabolism of [18F]fluorocholine in 9L glioma cells and 9L glioma-bearing fisher rats

  • Aditya Bansal
  • Wang Shuyan
  • Toshiko Hara
  • Robert A. Harris
  • Timothy R. DeGrado
Original Article



[18F]Fluorocholine ([18F]FCH) was developed as an analog of [11C]choline for tumor imaging; however, its metabolic handling remains ill defined. In this study, the metabolism of [18F]FCH is evaluated in cultured 9L glioma cells and Fisher 344 rats bearing 9L glioma tumors.


9L glioma cells were incubated with [18F]FCH and [14C]choline under normoxic and hypoxic (1% O2) conditions and analyzed for metabolic fate. [18F]FCH and [14C]choline kinetics and metabolism were studied in Fisher 344 rats bearing subcutaneous 9L tumors.


[18F]FCH and [14C]choline were similarly metabolized in 9L cells in both normoxic and hypoxic conditions over a 2-h incubation period. In normoxia, radioactivity was predominantly in phosphorylated form for both tracers after 5-min incubation. In hypoxia, the tracers remained mainly in nonmetabolized form at early timepoints (<20 min). Slow dephosphorylation of intracellular [18F]phosphofluorocholine (0.043–0.060 min−1) and [14C]phosphocholine (0.072–0.088 min−1) was evidenced via efflux measurements. In rat, both [18F]FCH and [14C]choline showed high renal and hepatic uptake. Blood clearance of both tracers was rapid with oxidative metabolites, [18F]fluorobetaine and [14C]betaine, representing the majority of radiolabel in plasma after 5 min postinjection. Oxidation (in liver) and lipid incorporation (in lung) were somewhat slower for [18F]FCH relative to [14C]choline. The majority of radiolabel in hypoxic subcutaneous tumor, as in hypoxic cultured 9L cells, was found as nonmetabolized [18F]FCH and [14C]choline.


[18F]FCH mimics choline uptake and metabolism by 9L glioma cells and tumors. However, subtle changes in biodistribution, oxidative metabolism, dephosphorylation, lipid incorporation, and renal excretion show moderate effects of the presence of the radiofluorine atom in [18F]FCH. The decrease in phosphorylation of exogenous choline by cancer cells should be considered in interpretation of positron emission tomography images in characteristically hypoxic tumors.


Choline Fluorocholine 189L glioma Rat Metabolism Hypoxia 



The work was funded in part by National Institutes of Health (RO1 CA108620, R01 HL-63371) and the Indiana Genomics Initiative Program of Indiana University School of Medicine (IUSM), a grant from the Lilly Endowment. The authors thank Dr. Frank A. Witzmann and Seokmin Hong in Department of Cellular and Integrative Physiology, IUSM for their help in liquid chromatography–mass spectrometry quantification of [19F]FCH.

Conflict of interest statement

No authors have affiliations that present conflicts of interest for this work.


  1. 1.
    Gillies RJ, Morse DL. In vivo magnetic resonance spectroscopy in cancer. Annu Rev Biomed Eng. 2005;7:287–26.CrossRefGoogle Scholar
  2. 2.
    DeGrado TR, Coleman RE, Wang S, Baldwin SW, Orr MD, Robertson CN, et al. Synthesis and evaluation of 18F-labeled choline as an oncologic tracer for positron emission tomography: initial findings in prostate cancer. Cancer Res. 2001;61(1):110–7.PubMedGoogle Scholar
  3. 3.
    Katz-Brull R, Seger D, Rivenson-Segal D, Rushkin E, Degani H. Metabolic markers of breast cancer: enhanced choline metabolism and reduced choline–ether–phospholipid synthesis. Cancer Res. 2002;62(7):1966–70.PubMedGoogle Scholar
  4. 4.
    Katz-Brull R, Degani H. Kinetics of choline transport and phosphorylation in human breast cancer cells: NMR application of the zero trans method. Anticancer Res. 1996;16(3B):1375–80.PubMedGoogle Scholar
  5. 5.
    Hara T, Kosaka N, Kishi H. Development of 18F-fluoroethylcholine for cancer imaging with PET: synthesis, biochemistry, and prostate cancer imaging. J Nucl Med. 2002;43(2):187–99.PubMedGoogle Scholar
  6. 6.
    Ramirez de Molina A, Rodriguez-Gonzalez A, Gutierrez R, Martinez-Pineiro L, Sanchez J, Bonilla F, et al. Overexpression of choline kinase is a frequent feature in human tumor-derived cell lines and in lung, prostate, and colorectal human cancers. Biochem Biophys Res Commun. 2002;296(3):580–3.PubMedCrossRefGoogle Scholar
  7. 7.
    Shinoura N, Nishijima M, Hara T, Haisa T, Yamamoto H, Fujii K, et al. Brain tumors: detection with C-11 choline PET. Radiology. 1997;202(2):497–503.PubMedGoogle Scholar
  8. 8.
    Hara T, Kosaka N, Shinoura N, Kondo T. PET imaging of brain tumor with [methyl-11C]choline. J Nucl Med. 1997;38(6):842–7.PubMedGoogle Scholar
  9. 9.
    Hara T, Kosaka N, Kishi H. PET imaging of prostate cancer using carbon-11-choline. J Nucl Med. 1998;39(6):990–5.PubMedGoogle Scholar
  10. 10.
    Hara M. Clinical studies on cefoperazone and polymyxin B for the treatment of infections in patients with hematological malignancies. Jpn J Antibiot. 1987;40(9):1639–43.PubMedGoogle Scholar
  11. 11.
    DeGrado TR, Baldwin SW, Wang S, Orr MD, Liao RP, Friedman HS, et al. Synthesis and evaluation of 18F-labeled choline analogs as oncologic PET tracers. J Nucl Med. 2001;42(12):1805–14.PubMedGoogle Scholar
  12. 12.
    Talbot JN, Gutman F, Fartoux L, Grange JD, Ganne N, Kerrou K, et al. PET/CT in patients with hepatocellular carcinoma using [18F]fluorocholine: preliminary comparison with [18F]FDG PET/CT. Eur J Nucl Med Mol Imaging 2006;33(11):1285–9. Jun 27.PubMedCrossRefGoogle Scholar
  13. 13.
    Kwee SA, Wei H, Sesterhenn I, Yun D, Coel MN. Localization of primary prostate cancer with dual-phase 18F-fluorocholine PET. J Nucl Med. 2006;47(2):262–9.PubMedGoogle Scholar
  14. 14.
    Heinisch M, Dirisamer A, Loidl W, Stoiber F, Gruy B, Haim S, et al. Positron emission tomography/computed tomography with F-18 fluorocholine for restaging of prostate cancer patients: meaningful at PSA<5 ng/ml? Mol Imaging Biol. 2006;8(1):43–8.PubMedCrossRefGoogle Scholar
  15. 15.
    Cimitan M, Bortolus R, Morassut S, Canzonieri V, Garbeglio A, Baresic T, et al. [18F] fluorocholine PET/CT imaging for the detection of recurrent prostate cancer at PSA relapse: experience in 100 consecutive patients. Eur J Nucl Med Mol Imaging 2006;33(12):1387–98. July 25.PubMedCrossRefGoogle Scholar
  16. 16.
    Schmid DT, John H, Zweifel R, Cservenyak T, Westera G, Goerres GW, et al. Fluorocholine PET/CT in patients with prostate cancer: initial experience. Radiology. 2005;235(2):623–8.PubMedCrossRefGoogle Scholar
  17. 17.
    Kwee SA, Coel MN, Lim J, Ko JP. Prostate cancer localization with fluorine-18 fluorocholine positron emission tomography. J Urol. 2005;173(1):252–5.PubMedCrossRefGoogle Scholar
  18. 18.
    Kwee SA, Coel MN, Lim J, Ko JP. Combined use of F-18 fluorocholine positron emission tomography and magnetic resonance spectroscopy for brain tumor evaluation. J Neuroimaging. 2004;14(3):285–9.PubMedCrossRefGoogle Scholar
  19. 19.
    Price DT, Coleman RE, Liao RP, Robertson CN, Polascik TJ, DeGrado TR. Comparison of [18F]fluorocholine and [18F]fluorodeoxyglucose for positron emission tomography of androgen dependent and androgen independent prostate cancer. J Urol. 2002;168(1):273–80.PubMedCrossRefGoogle Scholar
  20. 20.
    Roivainen A, Forsback S, Gronroos T, Lehikoinen P, Kahkonen M, Sutinen E, et al. Blood metabolism of [methyl-11C]choline; implications for in vivo imaging with positron emission tomography. Eur J Nucl Med. 2000;27(1):25–32.PubMedCrossRefGoogle Scholar
  21. 21.
    Jenkins WT, Evans SM, Koch CJ. Hypoxia and necrosis in rat 9L glioma and morris 7777 hepatoma tumors: comparative measurements using EF5 binding and the Eppendorf needle electrode. Int J Radiation Oncology Biol Phys. 2000;46(4):1005–17.Google Scholar
  22. 22.
    Liu L, Cash TP, Jones RG, Keith B, Thompson CB, Simon MC. Hypoxia-induced energy stress regulates mRNA translation and cell growth. Mol Cell. 2006;21:521–31.PubMedGoogle Scholar
  23. 23.
    Lamprecht W, Trautschold I. Determination of ATP by hexokinase and glucose-6-phosphate dehydrogenase. In: Bergmeyer HU, editor. Methods of Enzymatic Analysis. 2nd ed. New York: Academic; 1974. p. 2101–10.Google Scholar
  24. 24.
    Ishidate K, Nakazawa Y. Choline/ethanolamine kinase from rat kidney. Methods Enzymol. 1992;209:121–34.PubMedGoogle Scholar
  25. 25.
    Haubrich DR, Wang PF, Wedeking PW. Distribution and metabolism of intravenously administered choline[methyl- 3-H] and synthesis in vivo of acetylcholine in various tissues of guinea pigs. J Pharmacol Exp Ther. 1975;193(1):246–55.PubMedGoogle Scholar
  26. 26.
    Finkelstein JD, Martin JJ, Harris BJ, Kyle WE. Regulation of the betaine content of rat liver. Arch Biochem Biophys. 1982;218(1):169–73.PubMedCrossRefGoogle Scholar
  27. 27.
    Garcia-Perez A, Burg MB. Role of organic osmolytes in adaptation of renal cells to high osmolality. J Membr Biol. 1991;119:1–13.PubMedGoogle Scholar
  28. 28.
    Rooney SA, Young SL, Mendelson CR. Molecular and cellular processing of lung surfactant. Faseb J. 1994;8(12):957–67.PubMedGoogle Scholar
  29. 29.
    Pennington RJ, Worsfold M. Biosynthesis of lecithin by skeletal muscle. Biochim Biophys Acta. 1969;176(4):774–82.PubMedGoogle Scholar
  30. 30.
    Shamgar FA, Collins FD. Incorporation of ortho[32P]phosphate into phosphatidylcholines and phosphatidylethanolamines in rat skeletal muscle. Biochim Biophys Acta. 1975;409(1):104–15.PubMedGoogle Scholar
  31. 31.
    Hara T, Bansal A, DeGrado TR. Effect of hypoxia on uptake of [methyl-3H]choline, [1-14C]acetate and [18F]FDG in cultured prostate cancer cells. Nucl Med Biol. 2006;33(8):977–84. Nov.PubMedCrossRefGoogle Scholar
  32. 32.
    Sarri E, Garcia-Dorado D, Abellan A, Soler-Soler J. Effects of hypoxia, glucose deprivation and acidosis on phosphatidylcholine synthesis in HL-1 cardiomyocytes. CTP: phosphocholine cytidylyltransferase activity correlates with sarcolemmal disruption. Biochem J. 2006;394(1):325–34. Feb 15.PubMedGoogle Scholar
  33. 33.
    Jacobs RL, Lingrell S, Dyck JR, Vance DE. Inhibition of hepatic phosphatidylcholine synthesis by 5-aminoimidazole-4-carboxamide-1-beta-4-ribofuranoside is independent of AMP-activated protein kinase activation. J Biol Chem. 2007;282(7):4516–23. Feb 16.PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Aditya Bansal
    • 1
  • Wang Shuyan
    • 2
  • Toshiko Hara
    • 2
  • Robert A. Harris
    • 1
  • Timothy R. DeGrado
    • 1
    • 2
  1. 1.Department of Biochemistry and Molecular BiologyIndiana University School of MedicineIndianapolisUSA
  2. 2.Department of RadiologyIndiana University School of MedicineIndianapolisUSA

Personalised recommendations