Top

Cancer Chemotherapy and Pharmacology

Published in:

01-01-2011 | Original Article

The antioxidant ascorbic acid mobilizes nuclear copper leading to a prooxidant breakage of cellular DNA: implications for chemotherapeutic action against cancer

Authors: M. F. Ullah, H. Y. Khan, H. Zubair, U. Shamim, S. M. Hadi

Published in: Cancer Chemotherapy and Pharmacology | Issue 1/2011

Abstract

Purpose

Ascorbic acid is an essential micronutrient and is considered to have an antioxidant function in living systems. For the past several decades, ascorbic acid has been the subject of considerable interest as an anticancer agent. Several studies have shown that ascorbic acid is cytotoxic to a variety of cancer cells, whereas normal cells are relatively resistant to such cytotoxic action. In this study, we propose a putative molecular mechanism that accounts for the preferential cytotoxicity of ascorbic acid against cancer cells.

Methods

Standard and lysed version of alkaline single-cell gel electrophoresis (Comet assay); ferrous oxidation–xylenol orange (FOX) assay.

Results

We show that ascorbic acid acts as a prooxidant and leads to oxidative DNA breakage in lymphocytes and lymphocyte nuclei. Scavengers of reactive oxygen species were able to inhibit ascorbic acid-induced DNA breakage, suggesting the involvement of reactive oxygen species in this reaction. We further show that such DNA breakage is inhibited by both iron and copper chelators in cells, whereas in nuclei, similar inhibition was achieved only by copper chelators, indicating an important role of chromatin-bound copper in the prooxidant cellular DNA breakage by ascorbic acid.

Conclusion

We propose that the copper-dependent cellular redox status is an important element in the cytotoxic action of ascorbic acid against cancer cells. It is well established that cellular copper levels are considerably elevated in various malignancies. Therefore, cancer cells may be more subject to electron transfer between copper and ascorbate to generate reactive oxygen species. In light of these observations and those in literature, in this paper we explain that the preferential cytotoxicity of ascorbic acid against cancer cells is the result of elevated copper levels in such cells. Further, this study identifies nuclear copper as a novel molecular target for cytotoxic action of ascorbic acid, which has implications for its chemotherapeutic properties against cancer.

De Laurenzi V, Melino G, Savini I, Annicchiarico-Petruzelli M, Finnazi-agro A, Avigliano L (1995) Cell death by oxidative stress and ascorbic acid regeneration in human neuroectodermal cell lines. Eur J Cancer 31:463–466CrossRef

Riordan NH, Riordan HD, Meng X, Li Y, Jackson JA (1995) Intravenous ascorbate as tumor cytotoxic and tumor therapeutic agent. Med Hypothesis 44:207–213CrossRef

Park CH, Kim WS, Park C, Lee MH, Boo YC, Yoon SS et al (1999) Clinical disease suppression and reduction in acute myeloid leukemia and solid tumors by very high dose of L-ascorbic acid: a new concept and in search of molecular targets. Clin Cancer Res 5(Suppl 1):3784s

Kim WS, Lee MH, Chung CW, Kim K, Han SS, Park Q et al (2001) Clinical response in AML with manipulation of L-ascorbic acid (LAA) level in vivo to its extremes. Exp Hematol 29(Suppl 1):25

Chen Q, Espey MG, Krishna MC, Mitchell JB, Corpe CP, Buettner GR et al (2005) Pharmacologic ascorbic acid concentrations selectively kill cancer cells: action as a pro-drug to deliver hydrogen peroxide to tissues. Proc Natl Acad Sci USA 102:13604–13609CrossRefPubMed

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

Cameron E, Pauling L (1978) Supplemental ascorbate in the supportive treatment of cancer: reevaluation of prolongation of survival times in terminal human cancers. Proc Natl Acad Sci USA 75:4538–4542CrossRefPubMed

Creagen ET, Moertal CG, O’Fallon JR, Schutt AJ, O’Connell MJ, Rubin AJ et al (1979) Failure of high-dose Vitamin C (ascorbic acid) therapy to benefit patients with advanced cancer: a controlled trial. New England J Med 301:687–690CrossRef

Moertal CG, Fleming TR, Creagen ET, Rubin AJ, O’Connell MJ, Ames MM (1985) High-dose Vitamin C versus placebo in the treatment of patients with advanced cancer who have had no prior chemotherapy: a randomized double blind comparison. New England J Med 312:137–141CrossRef

10.

Omaya TS, Scala JH, Jacob RA (1986) Plasma ascorbic acid in adult males: effects of depletions and supplementation. J Am Coll Nutr 44:257–264

11.

Padayatty SJ, Sun H, Wang Y, Riordan HD, Hewit S, Katz A et al (2004) Vitamin C pharmacokinetics: implications for oral and intravenous use. Annuls Intern Med 140:533–538

12.

Chen Q, Espey MG, Sun AY, Pooput C, Kirk KL, Krishna MC, Khosh DB, Drisko J, Levine M (2008) Pharmacologic doses of ascorbate act as a prooxidant and decrease growth aggressive tumor xenografts in mice. Proc Natl Acad Sci USA 105:11105–11109CrossRefPubMed

13.

Halliwell B (2003) Oxidative stress in cell culture: an under appreciated problem? FEBS lett 540:3–6CrossRefPubMed

14.

Clement MV, Long LH, Ramalingam J, Halliwell B (2002) The cytotoxicity of dopamine may be an artifact of cell culture. J Neurochem 81:414–421CrossRefPubMed

15.

Shamim U, Hanif S, Ullah MF, Azmi AS et al (2008) Plant polyphenols mobilize nuclear copper in human peripheral lymphocytes leading to oxidatively generated DNA breakage: implications for an anticancer mechanism. Free Radic Res 42:764–772CrossRefPubMed

16.

Bhat SH, Azmi AS, Hanif S, Hadi SM (2006) Ascorbic acid mobilizes endogenous copper in human peripheral lymphocytes leading to oxidative DNA breakage: a putative mechanism for anticancer properties. IJBCB 38:2074–2081

17.

Smets KA (1994) Programmed cell death (apoptosis) and the response to anticancer drugs. Anticancer Drugs 5:3–9CrossRefPubMed

18.

Burkitt MJ, Milne L, Nicotera P, Orrenius S (1996) 1, 10-Phenanthroline stimulates internucleosomal DNA fragmentation in isolated rat liver nuclei by promoting redox activity of endogenous copper ions. Biochem J 313:163–169PubMed

19.

Linder MC (1991) Nutritional biochemistry and metabolism. Elsevier, New York

20.

Ebadi E, Swanson S (1998) The status of copper, zinc and metallothioneins in cancer patients. Prog Clin Biol Res 259:167–175

21.

Yoshida D, Ikada Y, Nakayama S (1993) Quantitative analysis of copper, zinc and copper/zinc ratios in selective human brain tumors. J Neuro-Oncol 16:109–115CrossRef

22.

Carpentieri U, Myers J, Thorpe L, Daeschner CW, Haggard ME (1986) Copper, zinc and iron in normal and leukemic lymphocytes in children. Cancer Res 46:981–984PubMed

23.

Pool-Zobel BL, Guigas C, Klein R, Neudecker CH, Renner HW, Schmezer P (1993) Assessment of genotoxic effects of lindane. Food Chem Toxicol 31:184–191

24.

Szeto YT, Collins AR, Benzie IFF (2002) Effects of dietary antioxidants on DNA damage in lysed cells using a modified comet assay procedure. Mutat Res 500:31–38PubMed

25.

Kasamatsu T, Kohda K, Kawazoe Y (1996) Comparison of chemically induced DNA breakage in cellular and subcellular systems using the comet assay. Mutat Res 369:1–6CrossRefPubMed

26.

Long LH, Clement MV, Halliwell B (2000) Artifacts in cell culture: rapid generation of hydrogen peroxide on addition of (−)-epigallocatechin, (−)-epigallocatechingallate, (+)-catechin and quercitin to commonly used cell culture media. Biochem Biophys Res Commun 273:50–53CrossRefPubMed

27.

Bhat R, Hadi SM (1994) DNA breakage by tannic acid–Cu(II): sequence specificity of the reaction and involvement of Cu(II). Mutat Res 313:39–48PubMed

28.

Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A, Kobayashi H et al (2000) Single cell gel electrophoresis/Comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen 35:206–221CrossRefPubMed

29.

Czene S, Tiback M, Harms-Ringdahl M (1997) pH-dependent DNA cleavage in permeabilized human fibroblasts. Biochem J 323:337–341PubMed

30.

Meneghini R, Martins EAL, Calderaro M (1993) Oxidative DNA damage and homeostasis of intracellular iron and copper ions. QUIMICA NOVA 16:377–380

31.

Toth I, Rogers JT, McPhee JA, Elliot SM, Abramson SL, Bridges KR (1995) Ascorbic acid enhances iron-induced ferritin translation in human leukemia and hepatoma cells. J Biol Chem 270:2846–2852CrossRefPubMed

32.

Scarpa M, Stevanato R, Vigilino P, Rigo A (1983) Superoxide ion as active intermediate in the autooxidation of ascorbate by molecular oxygen: effect of superoxide dismutase. J Biol Chem 258:6695–6697PubMed

33.

Gupte A, Mumper RG (2008) Elevated copper and oxidative stress in cancer cells as a target for cancer treatment. Cancer Treat Rev 35:32–46CrossRefPubMed

34.

Singh NP (1997) Sodium ascorbate induces DNA single-strand breaks in human cells in vitro. Mutat Res 375:195–203PubMed

35.

Kuo KW, Chen SF, Wu CC, Chen DR, Lee JH (2002) Serum and tissue trace elements in patients with breast cancer in Taiwan. Biol Trace Elem Res 89:1–11CrossRefPubMed

36.

Zuo XL, Chen JM, Zhou X, Li XL, Mei GY (2006) Levels of selenium, zinc, copper and antioxidant enzyme activity in patients with leukemia. Biol Trace Elem Res 114:41–54CrossRefPubMed

37.

Wolfe JT, Ross D, Cohen GM (1994) A role for metals and free radicals in the induction of apoptosis in thymocytes. FEBS Lett 352:59–62CrossRef

38.

Held KD, Sylvester FC, Hopcia KL, Biaglow JE (1996) Role of Fenton chemistry thiol-induced toxicity and apoptosis. Radiat Res 14:542–553CrossRef

39.

Pryor WA (1988) Why is hydroxyl radical the only radical that commonly binds to DNA? Hypothesis: it has rare combination of high electrophilicity, thermochemical reactivity and a mode of production near DNA. Free Rad Biol Med 4:219–233CrossRefPubMed

40.

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

41.

Devi GS, Prasad MH, Saraswathi I, Rao DN, Reddy PP (2000) Free radicals, antioxidant enzymes and lipid peroxidation in different types of leukemia. Clin Chim Acta 293:53–62CrossRefPubMed

42.

Schumacker PT (2006) Reactive oxygen species in cancer cells: live by the sword, die by the sword. Cancer Cell 10:175–176CrossRefPubMed

43.

Kong Q, Beel JA, Lillehei KO (2000) A threshold concept for cancer therapy. Med Hypothesis 55:29–35CrossRef

Title: The antioxidant ascorbic acid mobilizes nuclear copper leading to a prooxidant breakage of cellular DNA: implications for chemotherapeutic action against cancer
Authors: M. F. Ullah
H. Y. Khan
H. Zubair
U. Shamim
S. M. Hadi
Publication date: 01-01-2011
Publisher: Springer-Verlag
Published in: Cancer Chemotherapy and Pharmacology / Issue 1/2011
Print ISSN: 0344-5704
Electronic ISSN: 1432-0843
DOI: https://doi.org/10.1007/s00280-010-1290-4

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2011

Clinical characteristics and treatment outcomes of gastric cancer patients with isolated para-aortic lymph node involvement

Streptomycin alleviates irinotecan-induced delayed-onset diarrhea in rats by a mechanism other than inhibition of β-glucuronidase activity in intestinal lumen

In response to Dr. Han’s comments

Pioglitazone modulates tumor cell metabolism and proliferation in multicellular tumor spheroids

The small-molecule tyrosine kinase inhibitor nilotinib is a potent noncompetitive inhibitor of the SN-38 glucuronidation by human UGT1A1

Ultrasound-guided intratumoral administration of collagenase-2 improved liposome drug accumulation in solid tumor xenografts

Keynote webinar | Spotlight on antibody–drug conjugates in cancer