Chinese Science Bulletin

, Volume 44, Issue 12, pp 1101–1106 | Cite as

Apoptosis in suspension culture of tobacco cells induced by heat shock

  • Haoming Chen
  • Changhui Yan
  • Xiaofang Jiang
  • Yaoren Dai


The induction of apoptosis in suspension culture of tobacco cells by heat shock is reported for the first time. Heat treatment (48° for 4 h) of tobacco cells led to the appearance of typical hallmarks of apoptosis. It was demonstrated by DNA laddering analysis that the cells treated with heat shock at 48°C for 4 h had a serious degradation of nuclear DNA into multi-nucleosomal sizes, suggesting that heat shock activated endogenous nuclease which led to DNA cleavage at the linkage sites between the nucleosomes, but ladders were very faint for DNA from2 and 9 h heat-treated cells. The terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labelling (TUNEL) detection also showed that most of these treated cells (48°C for4 h) displayed positive reactions, indicating a serious DNA 3′-OH cleavage in their nuclei. Moreover, some other cytological changes in apoptotic cells, such as cell shrinkage, chromatin aggregation, nucleus collapse, have also been observed by 4′, 6′-diamidino-2-phenyl-indole (DAPI) staining.


apoptosis DNA ladder TUNEL Nicotiana tabacum 


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  1. 1.
    Kerr, J. F. R., Wylllie, A. H., Currie, A. H., Apoptosis: a basic biological phenomenon with wide ranging implications in tissue kinetics,Bri. J. Cancer., 1972, 26: 239.Google Scholar
  2. 2.
    McCabe, P. F., Levine, A., Meijer, P. al., A programmed cell death pathway activated in carrot cells cultured at low density,Plant J., 1997, 12(2): 267.CrossRefGoogle Scholar
  3. 3.
    Jiang, X. F., Zhu, H. Z., Zhou, al., Application of comet assay in plant protoplast apoptosis detection,Acta Botanica Sinica (in Chinese), 1998, 40(10): 928.Google Scholar
  4. 4.
    Poccia, F., Peselli, P., Vendetti, al., Heat shock protein expression on the membrane of T-cell undergoing apoptosis,Immnunology, 1996, 88: 6.CrossRefGoogle Scholar
  5. 5.
    Li, W. X., Chen, C. H., Ling, C. al., Apoptosis In heat induced cell killing the protective role of hsp-70 and the sensitization effect of the c-myc gene,Radh Res., 1996, 324.Google Scholar
  6. 6.
    Zhu, W. G., Aramaki, R., Cai, al., Promotion of heat-induced apoptosis in Fm3A cells by protease inhibitors,Biochem. Bioph. Res. Co., 1996, 225(3): 924.CrossRefGoogle Scholar
  7. 7.
    Vogel, P., Dux, E., Wiessner, C., Evidence of apoptosis in primary neuronal cultures after heat-shock.,Brain Res., 1997, 764(1-2): 205.PubMedCrossRefGoogle Scholar
  8. 8.
    Wang, H., Juan, L., Richard,. M. al., Apoptosis: a functional paradigm for programmed plant cell death induced by a host-selective phytotoxin and invoked during development,Plant Cell. 1996, 8: 375.PubMedCrossRefGoogle Scholar
  9. 9.
    Mitter, R., Lam, E., Identification, characterization, and purification of a tobacco endonuclease activity induced on hypersensitive response cell death,Plant Cell, 1995, 7: 1951.CrossRefGoogle Scholar
  10. 10.
    Harm, H. K., Thermotoleranee in mammalian cells protein denaturation and aggregation, and stress proteins,J. Cell Sci., 1993, 104: 11.Google Scholar
  11. 11.
    Mosser, D. D., Caron, A. W., Bourget, al., Role of the human hat-shock-protein Hsp70 in protection against stressinduced apoptosis,Mol. Cell Biol., 1997, 17(9): 5317.PubMedGoogle Scholar
  12. 12.
    Mehlen, P., Mehlen, A., Godet, al., Hap27 as a switch between differentiation and apoptosis in murine embryonic stem-cells,J. Biol. Chem., 1997, 272(50): 31657.PubMedCrossRefGoogle Scholar

Copyright information

© Science in China Press 1999

Authors and Affiliations

  • Haoming Chen
    • 1
  • Changhui Yan
    • 1
  • Xiaofang Jiang
    • 1
  • Yaoren Dai
    • 1
  1. 1.Department of BiologyTsinghua UniversityBeijingChina

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