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. 2003 Jan 15;369(Pt 2):239–248. doi: 10.1042/BJ20021021

Monitoring of apoptosis of HL60 cells by Fourier-transform infrared spectroscopy.

Fabio Gasparri 1, Marta Muzio 1
PMCID: PMC1223092  PMID: 12383054

Abstract

Fourier-transform infrared (FTIR) spectroscopy is a vibrational technique that gives information on the chemical composition of a sample, providing a "molecular fingerprint" of it. It is a powerful approach to study intact cells. The aim of the present study was to analyse and quantify apoptotic cells by using a FTIR approach based on attenuated total reflection (ATR). We incubated human HL60 leukaemic cells with camptothecin, a cytotoxic drug, and monitored apoptosis induction over a period of time. Several ATR-FTIR spectral changes occurred during the apoptotic process. In particular, we observed that the apoptotic index was inversely correlated with the spectral area in the region 1200-900 cm(-1), assigned to the absorption of nucleic acids. We therefore propose that ATR-FTIR spectral features may be used as a diagnostic marker of apoptotic cells.

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Selected References

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  1. Bertrand R., Solary E., Jenkins J., Pommier Y. Apoptosis and its modulation in human promyelocytic HL-60 cells treated with DNA topoisomerase I and II inhibitors. Exp Cell Res. 1993 Aug;207(2):388–397. doi: 10.1006/excr.1993.1206. [DOI] [PubMed] [Google Scholar]
  2. Boydston-White S., Gopen T., Houser S., Bargonetti J., Diem M. Infrared spectroscopy of human tissue. V. Infrared spectroscopic studies of myeloid leukemia (ML-1) cells at different phases of the cell cycle. Biospectroscopy. 1999;5(4):219–227. doi: 10.1002/(SICI)1520-6343(1999)5:4<219::AID-BSPY2>3.0.CO;2-O. [DOI] [PubMed] [Google Scholar]
  3. Cantrell D. A. Phosphoinositide 3-kinase signalling pathways. J Cell Sci. 2001 Apr;114(Pt 8):1439–1445. doi: 10.1242/jcs.114.8.1439. [DOI] [PubMed] [Google Scholar]
  4. Casiano C. A., Ochs R. L., Tan E. M. Distinct cleavage products of nuclear proteins in apoptosis and necrosis revealed by autoantibody probes. Cell Death Differ. 1998 Feb;5(2):183–190. doi: 10.1038/sj.cdd.4400336. [DOI] [PubMed] [Google Scholar]
  5. Cohen J. J. Apoptosis. Immunol Today. 1993 Mar;14(3):126–130. doi: 10.1016/0167-5699(93)90214-6. [DOI] [PubMed] [Google Scholar]
  6. Cohenford M. A., Rigas B. Cytologically normal cells from neoplastic cervical samples display extensive structural abnormalities on IR spectroscopy: implications for tumor biology. Proc Natl Acad Sci U S A. 1998 Dec 22;95(26):15327–15332. doi: 10.1073/pnas.95.26.15327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Del Bino G., Darzynkiewicz Z. Camptothecin, teniposide, or 4'-(9-acridinylamino)-3-methanesulfon-m-anisidide, but not mitoxantrone or doxorubicin, induces degradation of nuclear DNA in the S phase of HL-60 cells. Cancer Res. 1991 Feb 15;51(4):1165–1169. [PubMed] [Google Scholar]
  8. Dixit V. M. Role of ICE-proteases in apoptosis. Adv Exp Med Biol. 1996;406:113–117. doi: 10.1007/978-1-4899-0274-0_11. [DOI] [PubMed] [Google Scholar]
  9. Dolle R. E., Hoyer D., Prasad C. V., Schmidt S. J., Helaszek C. T., Miller R. E., Ator M. A. P1 aspartate-based peptide alpha-((2,6-dichlorobenzoyl)oxy)methyl ketones as potent time-dependent inhibitors of interleukin-1 beta-converting enzyme. J Med Chem. 1994 Mar 4;37(5):563–564. doi: 10.1021/jm00031a003. [DOI] [PubMed] [Google Scholar]
  10. Garrido C., Gurbuxani S., Ravagnan L., Kroemer G. Heat shock proteins: endogenous modulators of apoptotic cell death. Biochem Biophys Res Commun. 2001 Aug 24;286(3):433–442. doi: 10.1006/bbrc.2001.5427. [DOI] [PubMed] [Google Scholar]
  11. Gavrieli Y., Sherman Y., Ben-Sasson S. A. Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol. 1992 Nov;119(3):493–501. doi: 10.1083/jcb.119.3.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Goormaghtigh E., Cabiaux V., Ruysschaert J. M. Determination of soluble and membrane protein structure by Fourier transform infrared spectroscopy. III. Secondary structures. Subcell Biochem. 1994;23:405–450. doi: 10.1007/978-1-4615-1863-1_10. [DOI] [PubMed] [Google Scholar]
  13. Goormaghtigh E., Raussens V., Ruysschaert J. M. Attenuated total reflection infrared spectroscopy of proteins and lipids in biological membranes. Biochim Biophys Acta. 1999 Jul 6;1422(2):105–185. doi: 10.1016/s0304-4157(99)00004-0. [DOI] [PubMed] [Google Scholar]
  14. Goossens J. F., Hénichart J. P., Dassonneville L., Facompré M., Bailly C. Relation between intracellular acidification and camptothecin-induced apoptosis in leukemia cells. Eur J Pharm Sci. 2000 Apr;10(2):125–131. doi: 10.1016/s0928-0987(99)00091-3. [DOI] [PubMed] [Google Scholar]
  15. Herceg Z., Wang Z. Q. Functions of poly(ADP-ribose) polymerase (PARP) in DNA repair, genomic integrity and cell death. Mutat Res. 2001 Jun 2;477(1-2):97–110. doi: 10.1016/s0027-5107(01)00111-7. [DOI] [PubMed] [Google Scholar]
  16. Jamin N., Dumas P., Moncuit J., Fridman W. H., Teillaud J. L., Carr G. L., Williams G. P. Highly resolved chemical imaging of living cells by using synchrotron infrared microspectrometry. Proc Natl Acad Sci U S A. 1998 Apr 28;95(9):4837–4840. doi: 10.1073/pnas.95.9.4837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jesenberger Veronika, Jentsch Stefan. Deadly encounter: ubiquitin meets apoptosis. Nat Rev Mol Cell Biol. 2002 Feb;3(2):112–121. doi: 10.1038/nrm731. [DOI] [PubMed] [Google Scholar]
  18. Kaufmann S. H. Induction of endonucleolytic DNA cleavage in human acute myelogenous leukemia cells by etoposide, camptothecin, and other cytotoxic anticancer drugs: a cautionary note. Cancer Res. 1989 Nov 1;49(21):5870–5878. [PubMed] [Google Scholar]
  19. Liu K. Z., Jia L., Kelsey S. M., Newland A. C., Mantsch H. H. Quantitative determination of apoptosis on leukemia cells by infrared spectroscopy. Apoptosis. 2001 Aug;6(4):269–278. doi: 10.1023/a:1011383408381. [DOI] [PubMed] [Google Scholar]
  20. Martin S. J., Finucane D. M., Amarante-Mendes G. P., O'Brien G. A., Green D. R. Phosphatidylserine externalization during CD95-induced apoptosis of cells and cytoplasts requires ICE/CED-3 protease activity. J Biol Chem. 1996 Nov 15;271(46):28753–28756. doi: 10.1074/jbc.271.46.28753. [DOI] [PubMed] [Google Scholar]
  21. McDowell C. L., Papoutsakis E. T. Decreasing extracellular pH increases CD13 receptor surface content and alters the metabolism of HL60 cells cultured in stirred tank bioreactors. Biotechnol Prog. 1998 Jul-Aug;14(4):567–572. doi: 10.1021/bp980050w. [DOI] [PubMed] [Google Scholar]
  22. McDowell C. L., Papoutsakis E. T. Serum increases the CD13 receptor expression, reduces the transduction of fluid-mechanical forces, and alters the metabolism of HL60 cells cultured in agitated bioreactors. Biotechnol Bioeng. 1998 Oct 20;60(2):259–268. doi: 10.1002/(sici)1097-0290(19981020)60:2<259::aid-bit13>3.0.co;2-h. [DOI] [PubMed] [Google Scholar]
  23. Nicholson D. W., Ali A., Thornberry N. A., Vaillancourt J. P., Ding C. K., Gallant M., Gareau Y., Griffin P. R., Labelle M., Lazebnik Y. A. Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis. Nature. 1995 Jul 6;376(6535):37–43. doi: 10.1038/376037a0. [DOI] [PubMed] [Google Scholar]
  24. Piguet V., Wan L., Borel C., Mangasarian A., Demaurex N., Thomas G., Trono D. HIV-1 Nef protein binds to the cellular protein PACS-1 to downregulate class I major histocompatibility complexes. Nat Cell Biol. 2000 Mar;2(3):163–167. doi: 10.1038/35004038. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rigas B., Morgello S., Goldman I. S., Wong P. T. Human colorectal cancers display abnormal Fourier-transform infrared spectra. Proc Natl Acad Sci U S A. 1990 Oct;87(20):8140–8144. doi: 10.1073/pnas.87.20.8140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schultz C. P., Liu K. Z., Kerr P. D., Mantsch H. H. In situ infrared histopathology of keratinization in human oral/oropharyngeal squamous cell carcinoma. Oncol Res. 1998;10(5):277–286. [PubMed] [Google Scholar]
  27. Schultz C. P., Liu K., Johnston J. B., Mantsch H. H. Study of chronic lymphocytic leukemia cells by FT-IR spectroscopy and cluster analysis. Leuk Res. 1996 Aug;20(8):649–655. doi: 10.1016/0145-2126(96)00019-7. [DOI] [PubMed] [Google Scholar]
  28. Spiegel S. Sphingosine 1-phosphate: a prototype of a new class of second messengers. J Leukoc Biol. 1999 Mar;65(3):341–344. doi: 10.1002/jlb.65.3.341. [DOI] [PubMed] [Google Scholar]
  29. Surewicz W. K., Mantsch H. H. New insight into protein secondary structure from resolution-enhanced infrared spectra. Biochim Biophys Acta. 1988 Jan 29;952(2):115–130. doi: 10.1016/0167-4838(88)90107-0. [DOI] [PubMed] [Google Scholar]
  30. Tamm L. K., Tatulian S. A. Infrared spectroscopy of proteins and peptides in lipid bilayers. Q Rev Biophys. 1997 Nov;30(4):365–429. doi: 10.1017/s0033583597003375. [DOI] [PubMed] [Google Scholar]
  31. Tewari M., Quan L. T., O'Rourke K., Desnoyers S., Zeng Z., Beidler D. R., Poirier G. G., Salvesen G. S., Dixit V. M. Yama/CPP32 beta, a mammalian homolog of CED-3, is a CrmA-inhibitable protease that cleaves the death substrate poly(ADP-ribose) polymerase. Cell. 1995 Jun 2;81(5):801–809. doi: 10.1016/0092-8674(95)90541-3. [DOI] [PubMed] [Google Scholar]
  32. Willingham M. C. Cytochemical methods for the detection of apoptosis. J Histochem Cytochem. 1999 Sep;47(9):1101–1110. doi: 10.1177/002215549904700901. [DOI] [PubMed] [Google Scholar]
  33. Wyllie A. H. Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature. 1980 Apr 10;284(5756):555–556. doi: 10.1038/284555a0. [DOI] [PubMed] [Google Scholar]
  34. Zhou J., Wang Z., Sun S., Liu M., Zhang H. A rapid method for detecting conformational changes during differentiation and apoptosis of HL60 cells by Fourier-transform infrared spectroscopy. Biotechnol Appl Biochem. 2001 Apr;33(Pt 2):127–132. doi: 10.1042/ba20000074. [DOI] [PubMed] [Google Scholar]
  35. van Engeland M., Nieland L. J., Ramaekers F. C., Schutte B., Reutelingsperger C. P. Annexin V-affinity assay: a review on an apoptosis detection system based on phosphatidylserine exposure. Cytometry. 1998 Jan 1;31(1):1–9. doi: 10.1002/(sici)1097-0320(19980101)31:1<1::aid-cyto1>3.0.co;2-r. [DOI] [PubMed] [Google Scholar]

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