Skip to main content
SpringerLink
Log in

Ascorbate modulation of H2O2 and camptothecin-induced cell death in Jurkat cells

  • Original Article
  • Published:
Cancer Chemotherapy and Pharmacology Aims and scope Submit manuscript

Abstract

The effect of ascorbate on cell death was examined in Jurkat cells (human T-cell leukemia) by incubation with dehydroascorbate (DHA), which is rapidly taken up by cells and efficiently reduced to ascorbate. Apoptosis was evaluated by caspase-3 activity in cell extracts and flow cytometry of annexin V-labeled cells. In parallel, necrosis was estimated by the release of lactate dehydrogenase. Minor effects on cell death were observed when Jurkat cells were incubated with either DHA alone (100–1,000 μM) or a single dose of 10 μM H2O2. However, pre-incubation with DHA followed by exposure to H2O2 clearly stimulated both apoptosis and necrosis. In complete contrast, pre-incubation of cells with DHA significantly inhibited apoptosis, but did not affect necrosis, induced by the topoisomerase I inhibitor camptothecin. Our results indicate that intracellular ascorbate can modulate cell death in a manner which depends upon the nature of the apoptotic stimulus, which in turn has critical implications regarding the mechanism and potential application of ascorbate in cancer therapy.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

CPT:

20-S-camptothecin lactone

DHA:

Dehydroascorbate

LDH:

Lactate dehydrogenase

Ac-DEVD-AMC:

Ac-Asp-Glu-Val-Asp-amino-4-methylcoumarin

PS:

Phosphatidylserine

GSH:

Glutathione

DTT:

Dithiothreitol

References

  1. Agus DB, Vera JC, Golde DW (1999) Stromal cell oxidation: a mechanism by which tumors obtain vitamin C. Cancer Res 59:4555–4558

    CAS  PubMed  Google Scholar 

  2. Antunes F, Cadenas E (2001) Cellular titration of apoptosis with steady state concentrations of H2O2: submicromolar levels of H2O2 induce apoptosis through fenton chemistry independent of the cellular thiol state. Free Radic Biol Med 30:1008–1018

    Article  CAS  PubMed  Google Scholar 

  3. Asano K, Satoh K, Hosaka M, Arakawa H, Inagaki M, Hisamitsu T, Maeda M, Kochi M, Sakagami H (1999) Production of hydrogen peroxide in cancerous tissue by intravenous administration of sodium 5,6-benzylidene-l-ascorbate. Anticancer Res 19:229–236

    CAS  PubMed  Google Scholar 

  4. Bachleitner-Hofmann T, Gisslinger B, Grumbeck E, Gisslinger H (2001) Arsenic trioxide and ascorbic acid: synergy with potential implications for the treatment of acute myeloid leukaemia? Br J Haematol 112:783–786

    Article  CAS  PubMed  Google Scholar 

  5. Barbouti A, Doulias PT, Nousis L, Tenopoulou M, Galaris D (2002) DNA damage and apoptosis in hydrogen peroxide-exposed Jurkat cells: bolus addition versus continuous generation of H2O2. Free Radic Biol Med 33:691–702

    Article  CAS  PubMed  Google Scholar 

  6. Bijur GN, Briggs B, Hitchcock CL, Williams MV (1999) Ascorbic acid-dehydroascorbate induces cell cycle arrest at G2/M DNA damage checkpoint during oxidative stress. Environ Mol Mutagen 33:144–152

    Article  CAS  PubMed  Google Scholar 

  7. Cameron E, Campbell A (1974) The orthomolecular treatment of cancer. II. Clinical trial of high-dose ascorbic acid supplements in advanced human cancer. Chem Biol Interact 9:285–315

    Article  CAS  PubMed  Google Scholar 

  8. Casciari JJ, Riordan NH, Schmidt TL, Meng XL, Jackson JA, Riordan HD (2001) Cytotoxicity of ascorbate, lipoic acid, and other antioxidants in hollow fibre in vitro tumours. Br J Cancer 84:1544–1550

    Article  CAS  PubMed  Google Scholar 

  9. Chandra J, Samali A, Orrenius S (2000) Triggering and modulation of apoptosis by oxidative stress. Free Radic Biol Med 29:323–333

    Article  CAS  PubMed  Google Scholar 

  10. Chau YP, Shiah SG, Don MJ, Kuo ML (1998) Involvement of hydrogen peroxide in topoisomerase inhibitor beta-lapachone-induced apoptosis and differentiation in human leukemia cells. Free Radic Biol Med 24:660–670

    Article  CAS  PubMed  Google Scholar 

  11. Clement MV, Ramalingam J, Long LH, Halliwell B (2001) The in vitro cytotoxicity of ascorbate depends on the culture medium used to perform the assay and involves hydrogen peroxide. Antiox Red Sign 3:157–163

    Article  CAS  Google Scholar 

  12. Crott JW, Fenech M (1999) Effect of vitamin C supplementation on chromosome damage, apoptosis and necrosis ex vivo. Carcinogenesis 20:1035–1041

    Article  CAS  PubMed  Google Scholar 

  13. Guaiquil VH, Farber CM, Golde DW, Vera JC (1997) Efficient transport and accumulation of vitamin C in HL-60 cells depleted of glutathione. J Biol Chem 272:9915–9921

    Article  CAS  PubMed  Google Scholar 

  14. Guaiquil VH, Vera JC, Golde DW (2001) Mechanism of vitamin C inhibition of cell death induced by oxidative stress in glutathione-depleted HL-60 cells. J Biol Chem 276:40955–40961

    Article  CAS  PubMed  Google Scholar 

  15. Hampton MB, Orrenius S (1997) Dual regulation of caspase activity by hydrogen peroxide: implications for apoptosis. FEBS Lett 414:552–556

    Article  CAS  PubMed  Google Scholar 

  16. Hoffer LJ (2001) Proof versus plausibility: rules of engagement for the struggle to evaluate alternative cancer therapies. Can Med Assoc J 164:351–353

    CAS  Google Scholar 

  17. Hussein MA (2001) Arsenic trioxide—a new immunomodulatory agent in the management of multiple myeloma. Med Oncol 18:239–242

    Article  CAS  PubMed  Google Scholar 

  18. Kang JS, Cho DH, Kim YI, Hahm E, Yang YH, Kim D, Hur D, Park H, Bang S, Hwang YI, Lee WJ (2003) l-ascorbic acid (vitamin C) induces the apoptosis of B16 murine melanoma cells via a caspase-8-independent pathway. Cancer Immunol Immunother 52:693–698

    Article  CAS  PubMed  Google Scholar 

  19. Kaufmann SH (1998) Cell death induced by topoisomerase-targeted drugs: more questions than answers. Biochim Biophys Acta 1400:195–211

    Article  CAS  PubMed  Google Scholar 

  20. Koopman G, Reutelingsperger CP, Kuijten GA, Keehnen RM, Pals ST, van Oers MH (1994) Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood 84:1415–1420

    CAS  PubMed  Google Scholar 

  21. Lee YJ, Shacter E (2000) Hydrogen peroxide inhibits activation, not activity, of cellular caspase-3 in vivo. Free Radic Biol Med 29:684–692

    Article  CAS  PubMed  Google Scholar 

  22. Lee SH, Yoon YC, Jang YY, Song JH, Han ES, Lee CS (2001) Effect of iron and ascorbate on cyclosporine-induced oxidative damage of kidney mitochondria and microsomes. Pharmacol Res 43:161–171

    Article  CAS  PubMed  Google Scholar 

  23. Lenton KJ, Therriault H, Fulop T, Payette H, Wagner JR (1999) Glutathione and ascorbate are negatively correlated with oxidative DNA damage in human lymphocytes. Carcinogenesis 20:607–613

    CAS  PubMed  Google Scholar 

  24. Mizutani H, Tada-Oikawa S, Hiraku Y, Oikawa S, Kojima M, Kawanishi S (2002) Mechanism of apoptosis induced by a new topoisomerase inhibitor through the generation of hydrogen peroxide. J Biol Chem 277:30684–30689

    Article  CAS  PubMed  Google Scholar 

  25. Padayatty SJ, Levine M (2001) New insights into the physiology and pharmacology of vitamin C. Can Med Assoc J 164:353–355

    CAS  Google Scholar 

  26. Puskas F, Gergely P, Banki K, Perl A (2000) Stimulation of the pentose phosphate pathway and glutathione levels by dehydroascorbate, the oxidized form of vitamin C. Fed Am Soc Exp Biol J 14:1352–1361

    Article  CAS  Google Scholar 

  27. Ricci JE, Gottlieb RA, Green DR (2003) Caspase-mediated loss of mitochondrial function and generation of reactive oxygen species during apoptosis. J Cell Biol 160:65–75

    Article  CAS  PubMed  Google Scholar 

  28. Rose RC, Bode AM (1995) Analysis of water-soluble antioxidants by high-pressure liquid chromatography. Biochem J 306:101–105

    CAS  PubMed  Google Scholar 

  29. Sakagami H, Satoh K (1997) Modulating factors of radical intensity and cytotoxic activity of ascorbate. Anticancer Res 17:3513–3520

    CAS  PubMed  Google Scholar 

  30. Sakagami H, Satoh K, Hakeda Y, Kumegawa M (2000) Apoptosis-inducing activity of vitamin C and vitamin K. Cell Mol Biol 46:129–143

    CAS  Google Scholar 

  31. Sane AT, Bertrand R (1998) Distinct steps in DNA fragmentation pathway during camptothecin-induced apoptosis involved caspase-, benzyloxycarbonyl- and N-tosyl-l-phenylalanylchloromethyl ketone-sensitive activities. Cancer Res 58:3066–3072

    CAS  PubMed  Google Scholar 

  32. Schraufstatter IU, Hinshaw DB, Hyslop PA, Spragg RG, Cochrane CG (1985) Glutathione cycle activity and pyridine nucleotide levels in oxidant-induced injury of cells. J Clin Invest 76:1131–1139

    CAS  PubMed  Google Scholar 

  33. Schraufstatter IU, Hyslop PA, Hinshaw DB, Spragg RG, Sklar LA, Cochrane CG (1986) Hydrogen peroxide-induced injury of cells and its prevention by inhibitors of poly(ADP-ribose) polymerase. Proc Natl Acad Sci USA 83:4908–4912

    CAS  PubMed  Google Scholar 

  34. Shang F, Lu M, Dudek E, Reddan J, Taylor A (2003) Vitamin C and vitamin E restore the resistance of GSH-depleted lens cells to H2O2. Free Radic Biol Med 34:521–530

    Article  CAS  PubMed  Google Scholar 

  35. Shiah SG, Chuang SE, Chau YP, Shen SC, Kuo ML (1999) Activation of c-Jun NH2-terminal kinase and subsequent CPP32/Yama during topoisomerase inhibitor beta-lapachone induced apoptosis through an oxidation-dependent pathway. Cancer Res 59:391–398

    CAS  PubMed  Google Scholar 

  36. Song JH, Shin SH, Wang W, Ross GM (2001) Involvement of oxidative stress in ascorbate-induced proapoptotic death of PC12 cells. Exp Neurol 169:425–437

    Article  CAS  PubMed  Google Scholar 

  37. Spielholz C, Golde DW, Houghton AN, Nualart F, Vera JC (1997) Increased facilitated transport of dehydroascorbic acid without changes in sodium-dependent ascorbate transport in human melanoma cells. Cancer Res 57:2529–2537

    CAS  PubMed  Google Scholar 

  38. Szatrowski TP, Nathan CF (1991) Production of large amounts of hydrogen peroxide by human tumor cells. Cancer Res 51:794–798

    CAS  PubMed  Google Scholar 

  39. Taper HS, de Gerlache J, Lans M, Roberfroid M (1987) Non-toxic potentiation of cancer chemotherapy by combined C and K3 vitamin pre-treatment. Int J Cancer 40:575–579

    CAS  PubMed  Google Scholar 

  40. Verrax J, Cadrobbi J, Delvaux M, Jamison JM, Gilloteaux J, Summers JL, Taper HS, Calderon PB (2003) The association of vitamins C and K3 kills cancer cells mainly by autoschizis, a novel form of cell death. Basis for their potential use as coadjuvants in anticancer therapy. Eur J Med Chem 38:451–457

    Article  CAS  PubMed  Google Scholar 

  41. Younes M, Lechago LV, Somoano JR, Mosharaf M, Lechago J (1996) Wide expression of the human erythrocyte glucose transporter GLUT1 in human cancers. Cancer Res 56:1164–1167

    CAS  PubMed  Google Scholar 

  42. Younes M, Juarez D, Lechago LV, Lerner SP (2001) GLUT 1 expression in transitional cell carcinoma of the urinary bladder is associated with poor patient survival. Anticancer Res 21:575–578

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

A.T.S. was a recipient of a post-doctorate fellowship from le Fonds de Recherche en Santé du Québec. The authors would like to thank Dr Elliot Drobetsky for his helpful comments. This work was supported by a grant from the Cancer Research Society, Inc. Canada.

Author information

Authors and Affiliations

  1. Centre de recherche sur le vieillissement, Institut Universitaire de Gériatrie de Sherbrooke, Pavillion d’Youville, 1036, rue Belvédère Sud, Sherbrooke, QC, Canada, J1H 4C4

    Alain-Théophile Sané & J. Richard Wagner

  2. Unité de recherche pulmonaire, Centre Hospitalier Universitaire de Sherbrooke, Sherbrooke, QC, Canada, J1H 5N4

    André M. Cantin

  3. Département de médecine nucléaire et radiobiologie, Faculté de Médecine, Université de Sherbrooke, Sherbrooke, QC, Canada, J1H 5N4

    Benoit Paquette & J. Richard Wagner

Authors
  1. Alain-Théophile Sané
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. André M. Cantin
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Benoit Paquette
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. J. Richard Wagner
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to J. Richard Wagner.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sané, AT., Cantin, A.M., Paquette, B. et al. Ascorbate modulation of H2O2 and camptothecin-induced cell death in Jurkat cells. Cancer Chemother Pharmacol 54, 315–321 (2004). https://doi.org/10.1007/s00280-004-0828-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00280-004-0828-8

Keywords

Search

Navigation