Measurement of cell proliferation by labeling of DNA with stable isotope-labeled glucose: Studies <i>in vitro</i> , in animals, and in humans

  • Derek C. Macallan
    Department of Nutritional Sciences, University of California at Berkeley, Berkeley, CA 94720; and Department of Medicine, Division of Endocrinology and Metabolism, San Francisco General Hospital, University of California at San Francisco, San Francisco, CA 94110
  • Catherine A. Fullerton
    Department of Nutritional Sciences, University of California at Berkeley, Berkeley, CA 94720; and Department of Medicine, Division of Endocrinology and Metabolism, San Francisco General Hospital, University of California at San Francisco, San Francisco, CA 94110
  • Richard A. Neese
    Department of Nutritional Sciences, University of California at Berkeley, Berkeley, CA 94720; and Department of Medicine, Division of Endocrinology and Metabolism, San Francisco General Hospital, University of California at San Francisco, San Francisco, CA 94110
  • Katherine Haddock
    Department of Nutritional Sciences, University of California at Berkeley, Berkeley, CA 94720; and Department of Medicine, Division of Endocrinology and Metabolism, San Francisco General Hospital, University of California at San Francisco, San Francisco, CA 94110
  • Sunny S. Park
    Department of Nutritional Sciences, University of California at Berkeley, Berkeley, CA 94720; and Department of Medicine, Division of Endocrinology and Metabolism, San Francisco General Hospital, University of California at San Francisco, San Francisco, CA 94110
  • Marc K. Hellerstein
    Department of Nutritional Sciences, University of California at Berkeley, Berkeley, CA 94720; and Department of Medicine, Division of Endocrinology and Metabolism, San Francisco General Hospital, University of California at San Francisco, San Francisco, CA 94110

抄録

<jats:p> A method for measuring DNA synthesis and, thus, cell proliferation, <jats:italic>in vivo</jats:italic> is presented. The technique consists of administering [6,6- <jats:sup>2</jats:sup> H <jats:sub>2</jats:sub> ]Glc or [U- <jats:sup>13</jats:sup> C]Glc, isolating genomic DNA, hydrolyzing enzymatically to free deoxyribonucleosides, and derivatizing for GC-MS analysis of dA or dG isotopic enrichments, or both. Comparison of dA or dG to extracellular Glc enrichment (with a correction for intracellular dilution) reveals the fraction of newly synthesized DNA, by application of the precursor-product relationship. Thus, the technique differs from the widely used [ <jats:sup>3</jats:sup> H]thymidine or BrdUrd techniques in that the <jats:italic>de novo</jats:italic> nucleotide synthesis pathway, rather than the nucleoside salvage pathway, is used to label DNA; the deoxyribose rather than the base moiety is labeled; purine rather than pyrimidine deoxyribonucleosides are analyzed; and stable isotopes rather than radioisotopes are used. The method is applied here <jats:italic>in vitro</jats:italic> to the growth of HepG <jats:sub>2</jats:sub> and H <jats:sub>9</jats:sub> cells in culture; in animals to proliferation of intestinal epithelium, thymus, and liver; and in humans to granulocyte turnover in blood. In all instances, measured cell proliferation kinetics were consistent with expected or independently measured kinetics. The method has several advantages over previously available techniques for measuring cell turnover, involves no radioactivity or potentially toxic metabolites, and is suitable for use in humans. The availability of a reliable and safe method for measuring cell proliferation in humans opens up a number of fundamental questions to direct experimental testing, including basic problems related to cancer, AIDS, and other pathologic states. </jats:p>

収録刊行物

被引用文献 (3)*注記

もっと見る

キーワード

詳細情報

問題の指摘

ページトップへ