Ukr.Biochem.J. 2015; Volume 87, Issue 4, Jul-Aug, pp. 45-53


The sensitivity of cells with the various level of NAD(P)H:quinone oxidoreductase 1 to cytotoxic action of quinonimines and α-тоcopherol synthetic derivatives

G. V. Petrova1, А. V. Parshykov2

1Palladin Institute of Biochemistry, National Academy
of Sciences of Ukraine, Kyiv;
2Institute of Pharmacology and Toxocology, National Academy
of Medical Science of Ukraine, Kyiv

The effects of α-tocopherol with shortened to 6 carbon atoms side chain (α-Toc-C6), α-tocopherol succinate (α-TS) and quinonimine 2,6-dichlorophenolindophenol (DCPIP) on DT-diaphorase activity and viability of rat thymocytes, splenocytes and hepatocytes were investigated. It was shown that the lowest basal activity of the enzyme is inherent in splenocytes. In comparison to splenocytes, DT-diaphorase activity was 1.4 and 5 times higher in thymocytes and hepatocytes, respectively. It was found that the sensitivity of cells to the cytotoxic effect of DCPIP was inversely proportional to the basal level of DT-diaphorase activity and accompanied by its activation with subsequent inhibition at non-toxic and toxic concentrations, respectively. Hepatocytes were least sensitive to the cytotoxic effect of α-Toc-C6. In thymocytes and splenocytes α-Toc-C6 exerts inhibitory effects on DT-diaphorase, whereas in hepatocytes an increased activity of the enzyme was observed, which probably caused their high survival rate. Simultaneous induction of cytochrome P450 enzyme expression by α-Toc-C6 in hepatocytes is also possible. Cytotoxic effect of α-TS does not depend on the basal level of DT-diaphorase activity in cells, is not accompanied by its induction and it is most likely determined by the non-specific esterase activity.

Keywords: , , , , , ,


  1. Dinkova-Kostova AT, Talalay P. NAD(P)H:quinone acceptor oxidoreductase 1 (NQO1), a multifunctional antioxidant enzyme and exceptionally versatile cytoprotector. Arch Biochem Biophys. 2010 Sep 1;501(1):116-23. Review. PubMed, PubMedCentral, CrossRef
  2. Nioi P, Hayes JD. Contribution of NAD(P)H:quinone oxidoreductase 1 to protection against carcinogenesis, and regulation of its gene by the Nrf2 basic-region leucine zipper and the arylhydrocarbon receptor basic helix-loop-helix transcription factors. Mutat Res. 2004 Nov 2;555(1-2):149-71. Review. PubMed, CrossRef
  3. Zeekpudsa P, Kukongviriyapan V, Senggunprai L, Sripa B, Prawan A. Suppression of NAD(P)H-quinone oxidoreductase 1 enhanced the susceptibility of cholangiocarcinoma cells to chemotherapeutic agents. J Exp Clin Cancer Res. 2014 Jan 24;33:11. PubMed, PubMedCentral, CrossRef
  4. Cullen JJ, Hinkhouse MM, Grady M, Gaut AW, Liu J, Zhang YP, Weydert CJ, Domann FE, Oberley LW. Dicumarol inhibition of NADPH:quinone oxidoreductase induces growth inhibition of pancreatic cancer via a superoxide-mediated mechanism. Cancer Res. 2003 Sep 1;63(17):5513-20. PubMed
  5. Constantinou C, Papas A, Constantinou AI. Vitamin E and cancer: An insight into the anticancer activities of vitamin E isomers and analogs. Int J Cancer. 2008 Aug 15;123(4):739-52. Review. PubMed, CrossRef
  6. Petrova GV, Donchenko GV, Klimenko KP. Effects of α-tocopherol and its derivatives on the NAD(P)H:quinone oxidoreductase 1 level in rat thymocytes. Reports NASU. 2014;(4):156-161. (in Russian).
  7. Bellezza I, Grottelli S, Gatticchi L, Mierla AL, Minelli A. α-Tocopheryl succinate pre-treatment attenuates quinone toxicity in prostate cancer PC3 cells. Gene. 2014 Apr 10;539(1):1-7. PubMed, CrossRef
  8. Rooseboom M, Commandeur JN, Vermeulen NP. Enzyme-catalyzed activation of anticancer prodrugs. Pharmacol Rev. 2004 Mar;56(1):53-102. Review. PubMed, CrossRef
  9. Pat. 21527 UA, ICP A61K 31. 355, C07D 311. 72. A Technique for Obtaining of the 2,5,7,8-tetramethyl-2-(4-methyl-3-pentenyl)-6-chromanol acetate. Danevich O. I., Kirey Z. M., Andreichuk P.E., Kosenko М.V., Donchenko G.V., Klimenko K.P., Borutska Z.P., Borodina L.О., Svischuk О.А., Makovetsky V.P., Kuzmenko I.V. – Publ. 28.02.2000, Bul. N 1. (In Ukrainian).
  10. Lymphocytes. A practical approach. Edit by J. Klaus. Мoscw.: Мir, 1990. 393 p. (in Russian).
  11. Seglen PO. Prepаration of isolated rat liver cells: The enzymatic preparatıon of isolated intact parenchymal cells from rat liver. Methods Cell Biol. 1976;13:29-83. PubMed
  12. Prochaska HJ, Santamaria AB. Direct measurement of NAD(P)H:quinone reductase from cells cultured in microtiter wells: a screening assay for anticarcinogenic enzyme inducers. Anal Biochem. 1988 Mar;169(2):328-36. PubMed, CrossRef
  13. Bárta F, Levová K, Frei E, Schmeiser HH, Arlt VM, Stiborová M. The effect of aristolochic acid I on expression of NAD(P)H:quinone oxidoreductase in mice and rats–a comparative study. Mutat Res Genet Toxicol Environ Mutagen. 2014 Jul 1;768:1-7. PubMed, CrossRef
  14. Cabello CM, Bair WB 3rd, Bause AS, Wondrak GT. Antimelanoma activity of the redox dye DCPIP (2,6-dichlorophenolindophenol) is antagonized by NQO1. Biochem Pharmacol. 2009 Aug 15;78(4):344-54. PubMed, PubMedCentral, CrossRef
  15. Danson S, Ward TH, Butler J, Ranson M. DT-diaphorase: a target for new anticancer drugs. Cancer Treat Rev. 2004 Aug;30(5):437-49. Review. PubMed, CrossRef
  16. Siegel D, Ross D. Immunodetection of NAD(P)H:quinone oxidoreductase 1 (NQO1) in human tissues. Free Radic Biol Med. 2000 Aug;29(3-4):246-53. PubMed, CrossRef
  17. Petrova GV, Donchenko GV. Cytotoxicity of troglitazone, structural analogue of alpha-tocopherol is mediated by inhibition of NAD(P)H:quinone oxidoreductase. Ukr Biokhim Zhurn. 2009 Jul-Aug;81(4):105-11. Russian. PubMed
  18. Niki E, Traber MG. A history of vitamin E. Ann Nutr Metab. 2012;61(3):207-12. Review. PubMed, CrossRef
  19. Zuo ZY, Luo HL, Liu K, Jia HN, Zhang YW, Jiao LJ, Chang YF. Dietary vitamin E affects α-TTP mRNA levels in different tissues of the Tan sheep. Gene. 2014 May 10;541(1):1-7. PubMed, CrossRef
  20. Abe C, Uchida T, Ohta M, Ichikawa T, Yamashita K, Ikeda S. Cytochrome P450-dependent metabolism of vitamin E isoforms is a critical determinant of their tissue concentrations in rats. Lipids. 2007 Jul;42(7):637-45. PubMed, CrossRef
  21. Mustacich DJ, Leonard SW, Devereaux MW, Sokol RJ, Traber MG. Alpha-tocopherol regulation of hepatic cytochrome P450s and ABC transporters in rats. Free Radic Biol Med. 2006 Oct 1;41(7):1069-78. PubMed, CrossRef
  22. Landes N, Pfluger P, Kluth D, Birringer M, Rühl R, Böl GF, Glatt H, Brigelius-Flohé R. Vitamin E activates gene expression via the pregnane X receptor. Biochem Pharmacol. 2003 Jan 15;65(2):269-73. PubMed, CrossRef
  23. Tomasetti M, Santarelli L, Alleva R, Dong LF, Neuzil J. Redox-active and redox-silent compounds: synergistic therapeutics in cancer. Curr Med Chem. 2015;22(5):552-68. PubMed, CrossRef
  24. Zhao Y, Neuzil J, Wu K. Vitamin E analogues as mitochondria-targeting compounds: from the bench to the bedside? Mol Nutr Food Res. 2009 Jan;53(1):129-39. Review. PubMed, CrossRef
  25. Angulo-Molina A, Reyes-Leyva J, López-Malo A, Hernández J. The role of alpha tocopheryl succinate (α-TOS) as a potential anticancer agent. Nutr Cancer. 2014;66(2):167-76. Review. PubMed, CrossRef
  26. Zhang JG, Nicholls-Grzemski FA, Tirmenstein MA, Fariss MW. Vitamin E succinate protects hepatocytes against the toxic effect of reactive oxygen species generated at mitochondrial complexes I and III by alkylating agents. Chem Biol Interact. 2001 Dec 21;138(3):267-84. PubMed, CrossRef
  27. Farris MW, Fortuna MB, Everen CK,  Smith JD, Trent DF, Djuric Z. The selective antiproliferative effects of alpha-tocopheryl hemisuccinate and cholesteryl hemisuccinate on murine leukemia cells result from the action of the intact compounds. Cancer Res. 1994 Jul 1;54(13):3346-51. PubMed

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