Skip to main content
SpringerLink
Log in

Relationship between expression of a major apurinic/apyrimidinic endonuclease (APEX nuclease) and susceptibility to genotoxic agents in human glioma cell lines

  • Published:
Journal of Neuro-Oncology Aims and scope Submit manuscript

Abstract

The multifunctional DNA repair enzyme (APEX nuclease) having apurinic/apyrimidinic (AP) endonuclease, 3′-5′ exonuclease, DNA 3′ repair diesterase and DNA 3′-phosphatase activities is thought to be involved in repair of AP sites and single-strand breaks with 3′-blocked termini. To investigate the biological role of the enzyme, we studied the correlation between APEX AP endonuclease activity in several human glioma cell lines having various degree of its expression and cellular susceptibility to cytotoxic agents such as methyl methanesulfonate (MMS), 1-(4-amino-2-methyl-5-pyrimidinyl)methyl-3-(2-chloroethyl)-3-nitrosourea hy-drochloride (ACNU), cis-diamminedichloroplatinum(II) (CDDP), etoposide (VP-16), hydrogen peroxide (H2O2), hyperthermia and X-ray. The cell lines having lower APEX expression showed higher sensitivity to MMS and H2O2 which are known to induce AP sites and single strand breaks on DNA, respectively. The cellular susceptibility to the other agents tested was not significantly correlated to the APEX expression. The present results are thought to support the notion that APEX nuclease plays an important role on repair of AP sites and single-strand DNA breaks with 3′-blocked termini in mammalian cells.

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

Similar content being viewed by others

References

  1. Lindahl T: Instability and decay of the primary structure of DNA. Nature (Lond.) 362: 709–715, 1993

    Google Scholar 

  2. Sancar A, Sancar GB: DNA repair enzymes. Annu Rev Biochem 57: 29–67, 1988

    Google Scholar 

  3. Friedberg EC: DNA repair. W.H. Freeman, New York, 1985

    Google Scholar 

  4. Henner WD, Rodriguez LO, Hecht SM, Haseltine WA: γray induced deoxyribonucleic acid strand breaks. 3′ Glycolate termini. J Biol Chem 258: 711–713, 1983

    Google Scholar 

  5. Teebor GW, Boorstein RJ, Cadet J: The repairability of oxidative free radical mediated damage to DNA. A review. Int J Radiat Biol 54: 131–150, 1988

    Google Scholar 

  6. Loeb LA, Preston BD: Mutagenesis by apurinic/apyrimidinic sites. Annu Rev Genet 20: 201–230, 1986

    Google Scholar 

  7. Gentil A, Cabral-Neto JB, Mariage-Samson R, Margot A, Imbach JL, Rayner BR, Sarasin A: Mutagenicity of a unique apurinic/apyrimidinic site in mammalian cells. J Mol Biol 227: 981–984, 1992

    Google Scholar 

  8. Seki S, Oda T: An exonuclease possibly involved in the initiation of repair of bleomycin-damaged DNA in mouse ascites sarcoma cells. Carcinogenesis (Lond.) 9: 2239–2244, 1988

    Google Scholar 

  9. Seki S, Akiyama K, Watanabe S, Hatsushika M, Ikeda S, Tsutsui K: cDNA and deduced amino acid sequence of a mouse DNA repair enzyme (APEX nuclease) with significant homology toEscherichia coli exonuclease III. J Biol Chem 266: 20797–20802, 1991

    Google Scholar 

  10. Seki S, Ikeda S, Watanabe S, Hatsushika M, Tsutsui K, Akiyama K, Zhang B: A mouse DNA repair enzyme (APEX nuclease) having exonuclease and apurinic/apyrimidinic endonuclease activities: purification and characterization. Biochim Biophys Acta 1079: 57–64, 1991

    Google Scholar 

  11. Seki S, Hatsushika M, Watanabe S, Akiyama K, Nagao K, Tsutsui K: cDNA cloning, sequencing, expression and possible domain structure of human APEX nuclease homologous toEscherichia coli exonuclease III. Biochim Biophys Acta 1131: 287–299, 1992

    Google Scholar 

  12. Ono Y, Furuta T, Ohmoto T, Akiyama K, Seki S: Stable expression in rat glioma cells of sense and antisense nucleic acids to a human multifunctional DNA repair enzyme, APEX nuclease. Mutat Res 315: 55–63, 1994

    Google Scholar 

  13. Chomczynski P., Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159, 1987

    Google Scholar 

  14. Seki S, Akiyama K, Watanabe S, Tsutsui K: Activity gel and activity blotting methods for detecting DNA-modifying (repair) enzymes. J Chromatogra 618: 147–166, 1993

    Google Scholar 

  15. Walker MC, Masters JRW, Margison GP: O6-alkylguanine-DNA-alkyltransferase activity and nitrosourea sensitivity in human cancer cell lines. Br J Cancer 66: 840–843, 1992

    Google Scholar 

  16. Jordan JP, Hand CM, Markkowitz RS, Black P: Test for chemotherapeutic sensitivity of cerebral gliomas: use of colorimetric MTT assay. J Neuro-Oncology 14: 19–35, 1992

    Google Scholar 

  17. Nikkhah G, Tonn JC, Hoffmann O, Kraemer HP, Darling JL, Schachenmayr W, Schonmayr R: The MTT assay for chemosensitivity testing of human tumors of the central nervous system. J Neuro-Oncology 13: 13–24, 1992

    Google Scholar 

  18. Shiiki S, Fuchimoto S, Iwagaki H, Miyake M, Yasui Y, Orita K: Drug sensitivity test for interferon and tumor necrosis factorin vitro-comparison of dye uptake method and MTT assay. Biotherapy 2: 586–590, 1988

    Google Scholar 

  19. Mitsuhashi Y, Inaba M, Sugiyama Y, Kobayashi T:In vitro measurement of chemosensitivity of human small cell lung and gastric cancer cell lines toward cell cycle phase-nonspecific agents under the clinically equivalent area under the curve. Cancer 70: 2540–2546, 1992

    Google Scholar 

  20. Lin PS, Ho KC, Sung SJ, Gladding J: Effect of tumour necrosis factor, heat, and radiation on the viability and microfilament organization in cultured endothelial cells. Int J Hyperthermia 8: 667–677, 1992

    Google Scholar 

  21. Carmichael J, DeGraaf WG, Gazdar AF, Minna JD, Mitchell JB: Evaluation of a tetrazolium-based semiautomated colorimetric assay: Assessment of radiosensitivity. Cancer Res 47: 943–946, 1987

    Google Scholar 

  22. Hida T, Ueda R, Takahashi T, Watanabe H, Kato T, Suyama M, Sugiura R, Ariyoshi Y, Takahashi T: Chemosensitivity and radiosensitivity of small cell lung cancer cell lines studied by a newly developed 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) hybrid assay. Cancer Res 49: 4785–4790, 1989

    Google Scholar 

  23. Spearman G: The proof and measurement of association between two things. Am J Physiol 15: 72–101, 1994

    Google Scholar 

  24. Barnes DE, Lindahl T, Sedgwick B: DNA repair. Current Opinion in Cell Biology 5: 424–433, 1993

    Google Scholar 

  25. Robson CN, Hickson ID: Isolation of cDNA clones encoding a human apurinic/apyrimidinic endonuclease that corrects DNA repair and mutagenesis defects inE. coli xth (exonuclease III) mutants. Nucleic Acids Res 19: 5519–5523, 1991

    Google Scholar 

  26. Demple B, Herman T, Chen DS: Cloning and expression of APE, the cDNA encoding the major human apurinic endonuclease: Definition of a family of DNA repair enzymes. Proc Natl Acad Sci USA 88: 11450–11454, 1991

    Google Scholar 

  27. Xanthoudakis S, Miao G, Wang F, Pan Y-CE, Curran T: Redox activation of Fos-Jun DNA binding activity is mediated by a DNA repair enzyme. EMBO J 11: 3323–3335, 1992

    Google Scholar 

  28. Weiss B, Grossman L: Phosphodiesterases involved in DNA repair. Adv Enzymol 60: 1–34, 1987

    Google Scholar 

  29. Hoffmann MEE, Mello-Filho AC, Meneghini R: Correlation between cytotoxic effect of hydrogen peroxide and the yield of DNA strand breaks in cells of different species. Biochim Biophys Acta 781: 234–238, 1984

    Google Scholar 

  30. Imlay JA, Linn S: DNA damage and oxygen radical toxicity. Science (Washington DC) 240: 1302–1309, 1988

    Google Scholar 

  31. Halliwell B, Arouma OI: DNA damage by oxygen-derived species. Its mechanism and measurement in mammalian systems. FEBS Lett 281: 9–19, 1991

    Google Scholar 

  32. Hata H, Numata M, Tohda H, Yasui A, Oikawa A: Isolation of two chloroethylnitrosourea-sensitive Chinese hamster cell lines. Cancer Res 51: 195–198, 1991

    Google Scholar 

  33. Yajima N, Hochi S, Miyata N, Kishi T, Kawanishi G: DNA breaks and repair in the mouse leukemia L1210 cells exposed to three different types of interstrand DNA crosslinkers. Mutat Res 236: 43–50, 1990

    Google Scholar 

  34. Ishida R, Miki T, Narita T, Yui R, Sato M, Utsumi KR, Tanabe K, Andoh T: Inhibition of intracellular Topoisomerase II by antitumor bis(2,6-dioxopiperazine) derivatives: Mode of cell growth inhibition distinct from that of cleavable complex-forming type inhibitors. Cancer Res 51: 4909–4916, 1991

    Google Scholar 

  35. D'Arpa P, Lui LF: Topoisomerase-targeting antitumor drugs. Biochim Biophys Acta 989: 163–177, 1989

    Google Scholar 

  36. Cabral Neto JB, Gentil A, Caseira Cabral RE, Sarasin A: Mutation spectrum of heat-induced abasic sites on a singlestranded shuttle vector replicated in mammalian cells. J Biol Chem 267: 19718–19723, 1992

    Google Scholar 

  37. Yoshikawa T, Kokura S, Kondo M: The mechanism of the antitumor effect of hyperthermia: Role of free radical reaction in hyperthermia. Cancer Res Clin 2: 12–16, 1993

    Google Scholar 

  38. Lindquist S: The heat-shock response. Ann Rev Biochem 55: 1151–1191, 1986

    Google Scholar 

  39. Laurent G, Erickson LC, Sharkey NA, Kohn KW: DNA cross-linking and cytotoxicity induced by cis-diamminedi-chloroplatinum(II) in human normal and tumor cell lines. Cancer Res 41: 3347–3351, 1981

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Neurological Surgery, Okayama University Medical School, Okayama, Japan

    Yasuhiro Ono, Kengo Matsumoto, Tomohisa Furuta & Takashi Ohmoto

  2. Department of Molecular Biology, Institute of Cellular and Molecular Biology, Okayama University Medical School, Okayama, Japan

    Kosuke Akiyama & Shuji Seki

Authors
  1. Yasuhiro Ono
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kengo Matsumoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tomohisa Furuta
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Takashi Ohmoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kosuke Akiyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shuji Seki
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ono, Y., Matsumoto, K., Furuta, T. et al. Relationship between expression of a major apurinic/apyrimidinic endonuclease (APEX nuclease) and susceptibility to genotoxic agents in human glioma cell lines. J Neuro-Oncol 25, 183–192 (1995). https://doi.org/10.1007/BF01053151

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01053151

Key words

Search

Navigation