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Quantitative determination of apoptosis on leukemia cells by infrared spectroscopy

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Abstract

Quantitation of apoptotic cell death in vivo has become an important issue for patients with acute leukemia. We describe herein a new analytical method, based on infrared (IR) spectroscopy, to estimate the percentage of apoptotic leukemic cells in two different cell lines (CEM and K562), induced with etoposide (VP-16). As the percentage of apoptosis increases, the protein structure shifts from dominantly β-sheet to unordered (random coil), the overall lipid content increases and the amount of detectable DNA decreases. These changes can be directly related to the percentage of apoptosis as determined by two standard reference methods: flow cytometry and DNA ladder formation. The correlation between the significant IR spectral changes and the percentage of apoptotic leukemia cells in the two cell lines was optimal up to 24 h following etoposide treatment (r = 0.99 for CEM cells and r = 0.96 for K562 cells). Furthermore, IR spectroscopy is able to detect apoptotic changes in these cells already after 4 h treatment with VP-16, compared to flow cytometry which needs 6 h to observe significant changes. Our study suggests that IR spectroscopy may have potential clinical utility for the early, fast and reagent free assessment of chemotherapeutic efficacy in patients with leukemia.

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References

  1. Wylie AH. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 1980; 284: 555-556.

    Google Scholar 

  2. Blankenberg FG, Tait J, Ohtsuki K, Strauss HW. Apoptosis: The importance of nuclear medicine. Nucl Med Commun 2000; 21: 241-250.

    Google Scholar 

  3. Scott CE, Adebodun F. 13C-NMR investigation of protein synthesis during apoptosis in human leukemic cell lines. J Cell Physiol 1999; 181: 147-152.

    Google Scholar 

  4. Medina V, Edmonds B, Young GP, James R, Appleton S, Zalewski PD. Induction of caspase-3 protease activity and apoptosis by butyrate and trichostatin A (inhibitors of histone deacetylase): Dependence on protein synthesis and synergy with a mitochondrial/cytochrome c-dependent pathway. Cancer Res 1997; 57: 3697-3707.

    Google Scholar 

  5. Sweeley CC. Lipids and the modulation of cell function. Curr Topic Membr 1994; 40: 357-360.

    Google Scholar 

  6. Ichinose Y, Eguchi K, Migita K, et al. Apoptosis induction in synovial fibroblasts by ceramide: in vitro and in vivo effects. J Lab Clin Med 1998; 131: 410-416.

    Google Scholar 

  7. Willingham MC. Cytochemical methods for the detection of apoptosis. J Histochem Cytochem 1999; 47: 1101-1109.

    Google Scholar 

  8. Coelho D, Holl V, Weltin D, et al. Caspase-3-like activity determines the type of cell death following ionizing radiation in MOLT-4 human leukaemia cells. Br J Cancer 2000; 83: 642-649.

    Google Scholar 

  9. Kravtsov VD, Greer JP, Whitlock JA, Koury MJ. Use of the microculture kinetic assay of apoptosis to determine chemosensitivities of leukemias. Blood 1998; 92: 968-980.

    Google Scholar 

  10. Hannun YA. Apoptosis and the dilemma of cancer chemotherapy. Blood 1997; 89: 1845-1853.

    Google Scholar 

  11. Ozgen U, Savasan S, Buck S, Ravindranath Y. Comparison of DiOC(6)(3) uptake and annexin V labeling for quantification of apoptosis in leukemia cells and non-malignant T lymphocytes from children. Cytometry 2000; 42: 74-78.

    Google Scholar 

  12. Asselin BL, Ryan D, Frantz CN, Bernal SD, Leavitt P, Sallan SE, Cohen HJ. In vitro and in vivo killing of acute lymphoblastic leukemia cells by L-asparaginase. Cancer Res 1989; 49: 4363-4368.

    Google Scholar 

  13. Robertson LE, Chubb S, Meyn RE, Story M, Ford R, Hittelman WN, Plunkett W. Induction of apoptotic cell death in chronic lymphocytic leukemia by 2-chloro-2_-deoxyadenosine and 9-beta-D-arabinosyl-2-fluoroadenine. Blood 1993; 81: 143-150.

    Google Scholar 

  14. Schultz CP, Liu KZ, Kerr PD, Mantsch HH. In situ infrared histopathology of keratinization in human oral/oropharyngeal squamous cell carcinoma. Oncology Res 1998; 10: 277-286.

    Google Scholar 

  15. Schultz CP, Liu KZ, Salamon EA, Riese KT, Mantsch HH. Application of FT-IR microspectroscopy in diagnosing thyroid neoplasms. J Mol Struct 1999; 480-481: 369-377.

    Google Scholar 

  16. Jia L, Liu KZ, Newland AC, Mantsch HH, Kelsey SM. Increased mtDNA in Pgp positive leukaemic cells serves the energy requirement of the efflux pump but does not increase the malignant potential. Brit J Heamot 1999; 107: 861-869.

    Google Scholar 

  17. Liu KZ, Schultz CP, Mohammad RM, Johnston JB, Mantsch HH. Membrane alterations in a bryostatin 1 induced B-CLL cell line determinated by infrared spectroscopy. Leukemia 1999; 13: 1273-1280.

    Google Scholar 

  18. Liu KZ, Schultz CP, Johnston JB, Mantsch HH. Comparison of infrared spectra of CLL cells with their ex vivo sensitivity (MTT assay) to chlorambucil and cladribine. Leukemia Res 1997; 21: 1125-1133.

    Google Scholar 

  19. Schultz CP, Liu KZ, Johnston JB, Mantsch HH. Differentiation of leukemic from normal human lymphocytes by FT-IR spectroscopy and cluster analysis. Leukemia Res 1996; 20: 649-655.

    Google Scholar 

  20. Jia L, Kelsey SM, Grahn MF, Newland AC. Increased activity and sensitivity of mitochondrial respiratory enzymes to TNF?-induced inhibition is associated with cytotoxicity in drug-resistant leukemic cell lines. Blood 1996; 87: 2401-2410.

    Google Scholar 

  21. Jia L, Doumashkin RR, Allen PD, Gray AB, Newland AC, Kelsey SM. Inhibition of autophagy abrogates TNF?-induced apoptosis in human T-lymphoblastic leukaemic cells. Brit J Haematol 1997; 98: 673-685.

    Google Scholar 

  22. Jia L, Macey MG, Yin Y, Newland AC, Kelsey SM. Subcellular distribution and redistribution of Bcl-2 family proteins in human leukemic cells undergoing apoptosis. Blood 1999; 93: 2353-2359.

    Google Scholar 

  23. Griffiths PR, Pariente G. Trends in Analytical Chemistry. Elsevier Science, NY, Vol 5; 1986: 209-215.

    Google Scholar 

  24. Rigas R, Wong PTT. Human colon adenocarcinoma cell lines display infrared spectroscopic features of malignant colon tissues. Cancer Res 1992; 52: 84-88.

    Google Scholar 

  25. Surewicz WK, Mantsch HH. New insight into protein secondary structure from resolution-enhanced infrared spectra. Biochim Biophys Acta 1988; 952: 115-130.

    Google Scholar 

  26. Taillandier E, Liquier F. Infrared spectroscopy of DNA. Meth Enzymol 1992; 211: 307-335.

    Google Scholar 

  27. Le Gal JM, Manfait M, Theophanides. T. Applications of FT-IR spectroscopy in structural studies of cells and bacteria. J Mol Struct 1991; 242: 397-407.

    Google Scholar 

  28. Thornberry NA, Lasebnik Y. Caspases: Enemies within. Science 1998; 281: 1312-1316.

    Google Scholar 

  29. Benedetti E, Palatresi MP, Vergamini P, Papineschi F, Spremolla G. New possibilities of research in chronic lymphatic leukemia by means of Fourier transform-infrared spectroscopy-II. Leukemia Res 1985; 9: 1001-1008.

    Google Scholar 

  30. Gal JL, Morjani H, Manfait M. Ultrastructural appraisal of the multidrug resistance in K562 and LR73 cell lines from Fourier transform infrared spectroscopy. Cancer Res 1993; 53: 3681-3686.

    Google Scholar 

  31. Berkovic D, Fleer EA, Breass J, Pfortner J, Schleyer E, Hiddemann W. The influence of 1-beta-D-arabinofuranosylcytosine on the metabolism of phosphatidylcholine in human leukemic HL 60 and Raji cells. Leukemia 1997; 11: 2079-2086.

    Google Scholar 

  32. Haimovitz FA, Kolesnick RN, Fuks Z. Ceramide signaling in apoptosis. Br Med Bull 1997; 53: 539-553.

    Google Scholar 

  33. Maccarrone M, Nieuwenhuizen WE, Dullens HF, Catani MV, Melino G, Veldink GA, Vliegenthart JF, Finazzo-Agro A. Membrane modifications in human erythroleukemia K562 cells during induction of programmed cell death by transforming growth factor beta 1 or cisplatin. Eur J Biochem 1996; 241: 297-302.

    Google Scholar 

  34. Cohn RG, Mirkovich A, Dunlap B, Burton P, Chiu SH, Eugui E, Caulfield JP. Mycophenolic acid increases apoptosis, lysosomes and lipid droplets in human lymphoid and monocytic cell lines. Transplantation 1999; 68: 411-418.

    Google Scholar 

  35. Mohammad RM, Katato K, Almatchy VP, Wall N, Liu KZ, Schultz CP, Mantsch HH, Vartcrasian M, Al-Katib AM. Sequential treatment of human chronic lymphocytic leukemia with bryostatin 1 followed 2-chlorodeoxyadenosine: Preclinical studies. Clinical Cancer Res 1998; 4: 445-453.

    Google Scholar 

  36. Sasaki H, Suzuki T, Funaki N, et al. Immunohistochemistry of DNA fragmentation factor in human stomach and colon: its correlation to apoptosis Anticancer Res 1999; 19: 5277-5282.

    Google Scholar 

  37. Walker PR, Kokileva L, LeBlanc J, Sikorska M. Detection of the initial stages of DNA fragmentation in apoptosis. Biotechniques 1993; 15: 1032-1040.

    Google Scholar 

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Authors and Affiliations

  1. Institute for Biodiagnostics, National Research Council of Canada, Winnipeg, MB, Canada

    Kan-Zhi Liu

  2. Department of Haematology, St. Bartholomew's and The Royal London School of Medicine and Dentistry, Turner Street, London, UK

    Li Jia, S. M. Kelsey & A. C. Newland

  3. Institute for Biodiagnostics, National Research Council of Canada, Winnipeg, MB, Canada

    H. H. Mantsch

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  1. Kan-Zhi Liu
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  2. Li Jia
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  3. S. M. Kelsey
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  4. A. C. Newland
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  5. H. H. Mantsch
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Liu, KZ., Jia, L., Kelsey, S.M. et al. Quantitative determination of apoptosis on leukemia cells by infrared spectroscopy. Apoptosis 6, 269–278 (2001). https://doi.org/10.1023/A:1011383408381

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