Chinese Journal of Cancer Research

, Volume 9, Issue 1, pp 6–10 | Cite as

Analysis of apoptosis by DNA end labeling method (TDT) in leukemia cell lines HL-60 and U937

  • Li Ningli 
  • Shen Baihua 
  • Zheng Zexian 
  • Chou Guangyan 
Basic Investigations


Antitumor-drug-induced apoptosis in leukemia cell lines HL-60 and U937 was quantitatively analyzed with TDT (terminal deoxynuleotidyl transferase-mediated nick-end labeling) approach, which allows to label the ends of DNA broken strands in apoptotic cells by biotinlated dUTP which to avidin-FITC. In this way the apoptotic cells show fluorescent when the labeling cells were emitted by UV light microscope or laser-activated flow cell sorting at r=480. In our study, HL-60 and U937 cell lines were cocultured respectively with cis-diaminodichloroplatinum (CDDP), hydroxycamptothecinum (HCT) and vindesini sulfa (VCR) for 18 hours. By calculating percentages of apoptotic cells with TDT method, we were able to show that the two cell lines gave different sensitivity to the drugs. HL-60 showed high sensitivity to CDDP but U937 cells were more sensitive to other two drugs, HCT and VCR. Meanwhile we compared the results of obtained by DNA gel electrophoresis with that by TDT. We found that gel electrophoresis is not sensitive enough to reveal apoptosis since there was no ladder structure, a typical electrophoresis pattern for apoptosis, appeared until the apoptotic cells reached or over 13%. And we report in this paper as first time that three forms of apoptotic cells could be detected under fluorescent microscope, which we called as spot form and crescent form and assembling form in terms of distribution of light spots within cell nuclei. It seemed that the spot form was at an early stage of apoptosis and the crescent form represented a later stage of apoptosis.

Key words

Apoptosis Leukemia cell line Antitumor drug TDT 


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  1. 1.
    Duvall E, Wyllie AH. Death and the cell. Immunol Today 1986; 7:115.CrossRefGoogle Scholar
  2. 2.
    Wyllie AH. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 1980; 284:555.PubMedCrossRefGoogle Scholar
  3. 3.
    Gorczyca W, Gong JP, Darzynkiewicz A. Detection of DNA strand breaks in individual apoptosis cells by thein situ terminal deoxynucleotidyl transferase and nick translation assays. Cancer Res 1993; 53:1945.PubMedGoogle Scholar
  4. 4.
    Wesenlborg S, Janssen O, Kabelitz D. Induction of activation-driven death (apoptosis) in activated but not resting peripheral blood T cells. J Immunol 1993; 150:4338.Google Scholar
  5. 5.
    Kerr JFR. Shrinkage necrosis: A distinct mode of cellular death. J Pathol 1971; 105:13.PubMedCrossRefGoogle Scholar
  6. 6.
    Langlosis RG, Carrano AV, Gray JW, et al. Cytochemical studies of metaphase chromosomes by flow cytometry. Chromosoma 1980; 77:229.CrossRefGoogle Scholar
  7. 7.
    Sun XM, Snowden RT, Skilleter DN, et al. A flow-cytometric method for the separation and quantitation of normal and apoptosis thymocytes. Anal Biochem 1992; 204:351.PubMedCrossRefGoogle Scholar
  8. 8.
    1995;15:273.Google Scholar
  9. 9.
    Fehsel K, Kolb-Bachofen V, Kolb HJ. Analysis of TNFa-induced DNA strand breaks at the singe cell level. Am J Pathol 1991; 139:251.PubMedGoogle Scholar

Copyright information

© Chinese Journal of Cancer Research 1997

Authors and Affiliations

  • Li Ningli 
    • 1
  • Shen Baihua 
    • 1
  • Zheng Zexian 
    • 1
  • Chou Guangyan 
    • 1
  1. 1.Shanghai Institute of ImmunologyShanghai Second Medical UniversityShanghai

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