Measurement in vivo of proliferation rates of slow turnover cells by 2H2O labeling of the deoxyribose moiety of DNA
Measurement in vivo of proliferation rates of slow turnover cells by 2H2O labeling of the deoxyribose moiety of DNA
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DOI：
10.1073/pnas.232551499
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发表时间：
2002-11-26
影响因子：
11.1
通讯作者：
Hellerstein, MK
中科院分区：
文献类型：
--
作者：
Neese, RA;Misell, LM;Hellerstein, MK
我们在这里描述了一种方法，用于测量DNA复制，因此，细胞增殖的慢周转细胞，适用于人类。该技术是基于（H2O）-H-2掺入到嘌呤脱氧核糖核苷酸的脱氧核糖（dR）部分在分裂细胞。对于初始验证，向啮齿动物施用4%（H2O）-H-2的饮用水溶液。小鼠乳腺上皮细胞的增殖率为2.9%/d，在妊娠期间增加了5倍。给卵巢切除大鼠服用雌二醇颗粒（0-200 μ g）可增加乳腺上皮细胞增殖，根据剂量-反应关系，剂量高达100 μ g。同样，大鼠结肠上皮细胞的增殖刺激的剂量-反应方式由饮食胆酸。溴脱氧尿苷标记与（H2O）-H-2结果相关。然后测量慢更新细胞的增殖。从小鼠主动脉分离的血管平滑肌细胞分裂，半衰期为270-400天，（H2O）-H-2洗脱后的死亡值证实了这些缓慢的周转率。来自小鼠脂肪组织库的脂肪细胞富集部分的增殖率为每天1-1.5%的新细胞，而肥胖的自由采食的ob/ob小鼠表现出明显更高的增殖率和绝对增殖率。在人体中，通过每日摄入（H2O）-H-2，可实现体内水中稳定的长期（H2O）-H-2富集，且无毒性。来自完全翻转的血细胞（单核细胞或粒细胞）的标记dR显示出相对于体（H2O）-H-2富集的一致扩增因子（约3.5倍）。用（H2O）-H-2标记9周后新分裂的幼稚表型T细胞的分数为0.056（CD 4（+））和0.043（CD 8（+））（置换率
We describe here a method for measuring DNA replication and, thus, cell proliferation in slow turnover cells that is suitable for use in humans. The technique is based on the incorporation of (H2O)-H-2 into the deoxyribose (dR) moiety of purine deoxyribonucleotides in dividing cells. For initial validation, rodents were administered 4% (H2O)-H-2 in drinking water. The proliferation rate of mammary epithelial cells in mice was 2.9% per day and increased 5-fold during pregnancy. Administration of estradiol pellets (0-200 mug) to ovariectomized rats increased mammary epithelial cell proliferation, according to a dose-response relationship up to the 100 mug dose. Similarly, proliferation of colon epithelial cells was stimulated in a dose-response manner by dietary cholic acid in rats. Bromodeoxyuridine labeling correlated with the (H2O)-H-2 results. Proliferation of slow turnover cells was then measured. Vascular smooth muscle cells isolated from mouse aorta divided with a half-life in the range of 270-400 days and die-away values after (H2O)-H-2 wash-out confirmed these slow turnover rates. The proliferation rate of an adipocyte-enriched fraction from mouse adipose tissue depots was 1-1.5% new cells per day, whereas obese ad libitum-fed ob/ob mice exhibited markedly higher fractional and absolute proliferation rates. In humans, stable long-term (H2O)-H-2 enrichments in body water were achieved by daily (H2O)-H-2 intake, without toxicities. Labeled dR from fully turned-over blood cells (monocytes or granulocytes) exhibited a consistent amplification factor relative to body (H2O)-H-2 enrichment (approximate to3.5-fold). The fraction of newly divided naive-phenotype T cells after 9 weeks of labeling with (H2O)-H-2 was 0.056 (CD4(+)) and 0.043 (CD8(+)) (replacement rate