Tumor Biology

, Volume 33, Issue 4, pp 1021–1030 | Cite as

A combination of 2-deoxy-d-glucose and 6-aminonicotinamide induces cell cycle arrest and apoptosis selectively in irradiated human malignant cells

  • Richa Bhardwaj
  • Pradeep K. Sharma
  • S. P. S. Jadon
  • Rajeev Varshney
Research Article


Previously, we have shown that a combination of metabolic modifiers 2-deoxy-d-glucose (2-DG) and 6-aminonicotinamide (6-AN) results in oxidative stress mediated radiosensitization of malignant cells via mitochondrial dysfunction and non-coordinated expression of antioxidant defense, besides inhibition of repair and recovery. In the present study, our objective was to study, in a panel of human malignant cells of various origins (lung carcinoma, squamous carcinoma, oral carcinoma, and glioblastoma), if the inhibitory activity of combination (2-DG+6-AN+2Gy) against tumor growth could be considered a general phenomenon and to determine its effect on the cell cycle. The results revealed that combination (2-DG+6-AN+2Gy) treatment result in significant cell growth inhibition and induced ROS generation in all cancer cells studied. The anti-proliferative effect was related to the ability of combination (2-DG+6-AN+2Gy) to provoke growth inhibition at the G2/M arrest and apoptosis. Furthermore, combination (2-DG+6-AN+2Gy) induced G2/M arrest is closely correlated to decreased cyclin A, cyclin B1, and cdc2 levels.


Radiosensitization 2-Deoxy-d-glucose 6-Aminonicotinamide Oxidative stress G2/M arrest 







Reactive oxygen species


Pentose phosphate pathway




Fluorescein isothiocyanate


Propidium iodide


Phosphate buffered saline

Supplementary material

13277_2012_335_MOESM1_ESM.doc (90 kb)
ESM 1(DOC 90 kb)


  1. 1.
    Warburg O. On the origin of cancer cells. Science. 1956;132:309–14.CrossRefGoogle Scholar
  2. 2.
    Averill-Bates DA, Przybytkowski E. The role of glucose in cellular defenses against cytotoxicity of hydrogen peroxide in Chinese hamster ovary cells. Arch Biochem Biophys. 1994;312:52–8.PubMedCrossRefGoogle Scholar
  3. 3.
    Jelluma N, Yang X, Stokoe D, Evan GI, Dansen TB, Haas-Kogan DA. Glucose withdrawal induces oxidative stress followed by apoptosis in glioblastoma cells but not in normal human astrocytes. Mol Cancer Res. 2006;4:319–30.PubMedCrossRefGoogle Scholar
  4. 4.
    Lee YJ, Galoforo SS, Berns CM, Tong WP, Kim HRC, Corry PM. Glucose deprivation-induced cytotoxicity in drug resistant human breast carcinoma MCF-7/ADR cells: role of c-myc and bcl-2 in apoptotic cell death. J Cell Sci. 1997;110:681–6.PubMedGoogle Scholar
  5. 5.
    Singh D, Banerji AK, Dwarakanath BS, et al. Optimizing cancer radiotherapy with 2-deoxy-D-glucose: dose escalation studies in patients with cerebral glioblastoma multiforme. Strahlenther Onkol. 2005;181:507–14.PubMedCrossRefGoogle Scholar
  6. 6.
    Mohanti BK, Rath GK, Anantha N, et al. Improving cancer radiotherapy with 2-deoxy-D-glucose: phase I/II clinical trials on human cerebral gliomas. Int J Radiat Oncol Biol Phys. 1996;35:103–11.PubMedCrossRefGoogle Scholar
  7. 7.
    Jain V. Modifications of radiation responses by 2-deoxy-D-glucose in normal and cancer cells. Ind J Nucl Med. 1996;11:8–17.Google Scholar
  8. 8.
    Dwarakanath BS, Zolzer F, Chandna S, et al. Heterogeneity in 2-deoxy-D-glucose induced modifications in energetic and radiation responses of human tumor cell lines. Int J Radiat Oncol Biol Phys. 2001;51:1151–61.Google Scholar
  9. 9.
    Jha B, Pohlit W. Reversibility of inhibition of DNA double strand break repair by 2-deoxy-D-glucose in Ehrlich ascites tumour cells. Int J Radiat Biol. 1993;63:459–67.PubMedCrossRefGoogle Scholar
  10. 10.
    Weber G. Enzymology of cancer cells (first of two parts). New Eng J Med. 1977;296:486–92.PubMedCrossRefGoogle Scholar
  11. 11.
    Weber G. Enzymology of cancer cells (second of two parts). New Eng J Med. 1977;296:541–51.PubMedCrossRefGoogle Scholar
  12. 12.
    Varshney R, Dwarakanath BS, Jain V. Radiosensitization by 6-aminonicotinamide and 2-deoxy-D-glucose in human cancer cells. Int J Radiat Biol. 2005;81:397–408.PubMedCrossRefGoogle Scholar
  13. 13.
    Varshney R, Gupta S, Dwarakanath BS. Radiosensitization of Murine Ehrlich ascites tumor by a combination of 2-deoxy-D-glucose and 6-aminonicotinamide. Technol Cancer Res. 2004;3:659–63.Google Scholar
  14. 14.
    Sharma PK, Bhardwaj R, Dwarakanath BS, Varshney R. Metabolic oxidative stress induced by a combination of 2-DG and 6-AN enhances radiation damage selectively in malignant cells via non-coordinated expression of antioxidant enzymes. Cancer Lett. 2010;295:154–66.PubMedCrossRefGoogle Scholar
  15. 15.
    Bhardwaj R, Sharma PK, Jadon SPS, Varshney R. (2011) A combination of 2-deoxy-D-glucose and 6-aminonicotinamide induces oxidative stress mediated selective radiosensitization of malignant cells via mitochondrial dysfunction. Tumor Biol. 2011;32:951–64.CrossRefGoogle Scholar
  16. 16.
    Athar M, Chaudhury NK, Hussain ME, Varshney R. Hoechst 33342 enhances radiosensitization of malignant glioma cells via increase in mitochondrial ROS. Free Radic Res. 2010;44:936–49.PubMedCrossRefGoogle Scholar
  17. 17.
    Vermes I, Haanen C, Steffens-Nakken H, Reutelingsoerger C. A novel assay for apoptosis: flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labeled detection. J Immunol Methods. 1995;184:39–51.PubMedCrossRefGoogle Scholar
  18. 18.
    Chang JE, Khuntia D, Robins HI, Mehta MP. Radiotherapy and radiosensitizers in the treatment of glioblastoma multiforme. Clin Adv Hematol Oncol. 2007;5:894–915.PubMedGoogle Scholar
  19. 19.
    Singh SV, Srivastava SK, Choi S, et al. Sulphoraphane-induced cell death in human prostate cancer cells is initiated by reactive oxygen species. J Biol Chem. 2005;280:19911–24.PubMedCrossRefGoogle Scholar
  20. 20.
    Wu XJ, Hua X. Targeting ROS: selective killing of cancer cells by a cruciferous vegetable derived pro-oxidant compound. Cancer Biol Ther. 2007;6:646–7.PubMedCrossRefGoogle Scholar
  21. 21.
    Choi WY, Choi BT, Lee WH, Choi YH. Sulforaphane generates reactive oxygen species leading to mitochondrial perturbation for apoptosis in human leukemia U937 cells. Biomed Pharmacother. 2008;62:637–44.PubMedCrossRefGoogle Scholar
  22. 22.
    Zhang R, Humphreys I, Sahu RP, Shi Y, Srivastava SK. In vitro and in vivo induction of apoptosis by capsaicin in pancreatic cancer cells is mediated through ROS generation and mitochondrial death pathway. Apoptosis. 2008;13:1465–78.PubMedCrossRefGoogle Scholar
  23. 23.
    Li M, Zhao L, Liu J, et al. Hydrogen peroxide induces G2 cell cycle arrest and inhibits cell proliferation in osteoblasts. Anat Rec. 2009;292:1107–13.CrossRefGoogle Scholar
  24. 24.
    Sahu RP, Zhang R, Batra S, Shi Y, Srivastava SK. Benzyl isothiocyanate-mediated generation of reactive oxygen species causes cell cycle arrest and induces apoptosis via activation of MAPK in human pancreatic cancer cells. Carcinogenesis. 2009;30:1744–53.PubMedCrossRefGoogle Scholar
  25. 25.
    Burhans WC, Heintz NH. The cell cycle is a redox cycle: linking phase-specific targets to cell fate. Free Radic Biol Med. 2009;47:1282–93.PubMedCrossRefGoogle Scholar
  26. 26.
    Tyagi AK, Singh RP, Agarwal C, Chan DC, Agarwal R. Silibinin strongly synergizes human prostate carcinoma DU145 cells to doxorubicin-induced growth inhibition, G2-M arrest, and apoptosis. Clin Cancer Res. 2002;8:3512–9.PubMedGoogle Scholar
  27. 27.
    Singh RP, Dhanalakshmi S, Agarwal R. Phytochemicals as cell cycle modulators a less toxic approach in halting human cancers. Cell Cycle. 2002;1:156–61.PubMedCrossRefGoogle Scholar
  28. 28.
    King KL, Cidlowski JA. Cell cycle regulation and apoptosis. Annu Rev Physiol. 1998;60:601–17.PubMedCrossRefGoogle Scholar
  29. 29.
    Evan GI, Brown L, Whyte M, Harrington E. Apoptosis and the cell cycle. Curr Opin Cell Biol. 1995;7:825–34.PubMedCrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2012

Authors and Affiliations

  • Richa Bhardwaj
    • 1
  • Pradeep K. Sharma
    • 1
    • 2
  • S. P. S. Jadon
    • 3
  • Rajeev Varshney
    • 1
    • 4
  1. 1.Institute of Nuclear Medicine and Allied Sciences, DRDODelhiIndia
  2. 2.School of BiotechnologyGuru Gobind Singh Indraprastha UniversityDwarkaIndia
  3. 3.Department of ChemistryShri Varshneya CollegeAligarhIndia
  4. 4.DRDO Bhawan, Defence Research and Development Organisation, Ministry of DefenceGovernment of IndiaNew DelhiIndia

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