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Adaphostin cytoxicity in glioblastoma cells is ROS-dependent and is accompanied by upregulation of heme oxygenase-1

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Abstract

Purpose

To delineate a role for reactive oxygen species (ROS) induction in adaphostin-induced apoptosis in glioblastoma cells.

Methods

Three glioblastoma cell lines with different sensitivities to adaphostin were characterized for sensitivity to an oxidant, tert-butyl hydroperoxide. The degree and duration of the ROS levels was assessed in the three cell lines after adaphostin exposure. Antioxidant protein levels were evaluated by Western blotting.

Results

Of the three glioblastoma cell lines, the U87 cells were least sensitive to adaphostin. These cells were also least sensitive to tert-butyl hydroperoxide, indicating that sensitivity to a direct oxidant stress mirrors the cells’ adaphostin sensitivities. In addition, the antioxidant N-acetylcysteine, (NAC) was protective against adaphostin-induced apoptosis. Direct measurement of intracellular peroxides showed a transient increase in the two less sensitive cell lines (U87 and LN18) which diminishes by 24 h. In contrast, U251 cells, which are most sensitive to adaphostin, display a sustained increase in the ROS levels. After the initial increase in intracellular peroxides, the heat shock protein and antioxidant heme oxygenase-1 (HO-1) was upregulated. Levels of other antioxidant proteins, such as catalase and thioredoxin, however, were not altered by adaphostin in glioblastoma cell lines. NAC attenuated HO-1 upregulation, confirming the time course analysis.

Conclusions

These results suggest a primary role for ROS in adaphostin-induced apoptosis in glioblastoma. Our data indicate that the duration of intracellular ROS levels is a key factor in mediating sensitivity to adaphostin. Furthermore, upregulation of HO-1 is a novel molecular marker of adaphostin’s action. The kinetics with which adaphostin upregulates HO-1 correlates with sensitivity to the drug. Taken together, our data indicate that a cell’s ability to cope with ROS dictates sensitivity to adaphostin and conceivably other chemotherapies that cause redox perturbations.

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References

  1. Avramis IA, Christodoulopoulos G, Suzuki A, Laug WE, Gonzalez-Gomez I, McNamara G, Sausville EA, Avramis VI (2002) In vitro and in vivo evaluations of the tyrosine kinase inhibitor NSC 680410 against human leukemia and glioblastoma cell lines. Cancer Chemother Pharmacol 50:479–489

    Article  PubMed  CAS  Google Scholar 

  2. Burton EC, Prados MD (2000) Malignant gliomas. Curr Treat Options Oncol 1:459–468

    Article  PubMed  CAS  Google Scholar 

  3. Cao C, Leng Y, Liu X, Yi Y, Li P, Kufe D (2003) Catalase is regulated by ubiquitination and proteosomal degradation. Role of the c-Abl and Arg tyrosine kinases. Biochemistry 42:10348–10353

    Article  PubMed  CAS  Google Scholar 

  4. Chandra J, Hackbarth J, Le S, Loegering D, Bone N, Bruzek LM, Narayanan VL, Adjei AA, Kay NE, Tefferi A, Karp JE, Sausville EA, Kaufmann SH (2003) Involvement of reactive oxygen species in adaphostin-induced cytotoxicity in human leukemia cells. Blood 102:4512–4519

    Article  PubMed  CAS  Google Scholar 

  5. Chandra J, Tracy J, Loegering D, Flatten K, Verstovsek S, Beran M, Gorre M, Estrov Z, Donato N, Talpaz M, Sawyers C, Bhalla K, Karp J, Sausville E, Kaufmann SH (2005) Adaphostin-induced oxidative stress overcomes bcr/abl mutation-dependent and -independent imatinib resistance. Blood

  6. Dasmahapatra G, Nguyen TK, Dent P, Grant S (2006) Adaphostin and bortezomib induce oxidative injury and apoptosis in imatinib mesylate-resistant hematopoietic cells expressing mutant forms of Bcr/Abl. Leuk Res

  7. Hara E, Takahashi K, Tominaga T, Kumabe T, Kayama T, Suzuki H, Fujita H, Yoshimoto T, Shirato K, Shibahara S (1996) Expression of heme oxygenase and inducible nitric oxide synthase mRNA in human brain tumors. Biochem Biophys Res Commun 224:153–158

    Article  PubMed  CAS  Google Scholar 

  8. Hirota K, Murata M, Sachi Y, Nakamura H, Takeuchi J, Mori K, Yodoi J (1999) Distinct roles of thioredoxin in the cytoplasm and in the nucleus. A two-step mechanism of redox regulation of transcription factor NF-kappaB. J Biol Chem 274:27891–27897

    Article  PubMed  CAS  Google Scholar 

  9. Hose C, Kaur G, Sausville EA, Monks A (2005) Transcriptional profiling identifies altered intracellular labile iron homeostasis as a contributing factor to the toxicity of adaphostin: decreased vascular endothelial growth factor secretion is independent of hypoxia-inducible factor-1 regulation. Clin Cancer Res 11:6370–6381

    Article  PubMed  CAS  Google Scholar 

  10. Hovinga KE, Stalpers LJ, van Bree C, Donker M, Verhoeff JJ, Rodermond HM, Bosch DA, van Furth WR (2005) Radiation-enhanced vascular endothelial growth factor (VEGF) secretion in glioblastoma multiforme cell lines—a clue to radioresistance? J Neurooncol 74:99–103

    Article  PubMed  CAS  Google Scholar 

  11. Kawai H, Nie L, Yuan ZM (2002) Inactivation of NF-kappaB-dependent cell survival, a novel mechanism for the proapoptotic function of c-Abl. Mol Cell Biol 22:6079–6088

    Article  PubMed  CAS  Google Scholar 

  12. Lee HC, Kim DW, Jung KY, Park IC, Park MJ, Kim MS, Woo SH, Rhee CH, Yoo H, Lee SH, Hong SI (2004) Increased expression of antioxidant enzymes in radioresistant variant from U251 human glioblastoma cell line. Int J Mol Med 13:883–887

    PubMed  CAS  Google Scholar 

  13. Levitzki A, Gazit A (1995) Tyrosine kinase inhibition: an approach to drug development. Science 267:1782–1788

    Article  PubMed  CAS  Google Scholar 

  14. Li J, Lee JM, Johnson JA (2002) Microarray analysis reveals an antioxidant responsive element-driven gene set involved in conferring protection from an oxidative stress-induced apoptosis in IMR-32 cells. J Biol Chem 277:388–394

    Article  PubMed  CAS  Google Scholar 

  15. Maines MD (2005) New insights into biliverdin reductase functions: linking heme metabolism to cell signaling. Physiology (Bethesda) 20:382–389

    CAS  Google Scholar 

  16. Mayerhofer M, Florian S, Krauth MT, Aichberger KJ, Bilban M, Marculescu R, Printz D, Fritsch G, Wagner O, Selzer E, Sperr WR, Valent P, Sillaber C (2004) Identification of heme oxygenase-1 as a novel BCR/ABL-dependent survival factor in chronic myeloid leukemia. Cancer Res 64:3148–3154

    Article  PubMed  CAS  Google Scholar 

  17. Miralem T, Hu Z, Torno MD, Lelli KM, Maines MD (2005) Small interference RNA-mediated gene silencing of human biliverdin reductase, but not that of heme oxygenase-1, attenuates arsenite-mediated induction of the oxygenase and increases apoptosis in 293A kidney cells. J Biol Chem 280:17084–17092

    Article  PubMed  CAS  Google Scholar 

  18. Mow BM, Chandra J, Svingen PA, Hallgren CG, Weisberg E, Kottke TJ, Narayanan VL, Litzow MR, Griffin JD, Sausville EA, Tefferi A, Kaufmann SH (2002) Effects of the Bcr/abl kinase inhibitors STI571 and adaphostin (NSC 680410) on chronic myelogenous leukemia cells in vitro. Blood 99:664–671

    Article  PubMed  CAS  Google Scholar 

  19. Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C (1991) A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 139:271–279

    Article  PubMed  CAS  Google Scholar 

  20. Ohgaki H, Kleihues P (2005) Population-based studies on incidence, survival rates, and genetic alterations in astrocytic and oligodendroglial gliomas. J Neuropathol Exp Neurol 64:479–489

    PubMed  CAS  Google Scholar 

  21. Riley PA (1994) Free radicals in biology: oxidative stress and the effects of ionizing radiation. Int J Radiat Biol 65:27–33

    Article  PubMed  CAS  Google Scholar 

  22. Robinson JP, Bruner LH, Bassoe CF, Hudson JL, Ward PA, Phan SH (1988) Measurement of intracellular fluorescence of human monocytes relative to oxidative metabolism. J Leukoc Biol 43:304–310

    PubMed  CAS  Google Scholar 

  23. Sadowska AM, Manuel YKB, De Backer WA (2006) Antioxidant and anti-inflammatory efficacy of NAC in the treatment of COPD: discordant in vitro and in vivo dose-effects: a review. Pulm Pharmacol Ther

  24. Shanafelt TD, Lee YK, Bone ND, Strege AK, Narayanan VL, Sausville EA, Geyer SM, Kaufmann SH, Kay NE (2005) Adaphostin-induced apoptosis in CLL B cells is associated with induction of oxidative stress and exhibits synergy with fludarabine. Blood 105:2099–2106

    Article  PubMed  CAS  Google Scholar 

  25. Svingen PA, Tefferi A, Kottke TJ, Kaur G, Narayanan VL, Sausville EA, Kaufmann SH (2000) Effects of the bcr/abl kinase inhibitors AG957 and NSC 680410 on chronic myelogenous leukemia cells in vitro. Clin Cancer Res 6:237–249

    PubMed  CAS  Google Scholar 

  26. Yu C, Rahmani M, Almenara J, Sausville EA, Dent P, Grant S (2004) Induction of apoptosis in human leukemia cells by the tyrosine kinase inhibitor adaphostin proceeds through a RAF-1/MEK/ERK- and AKT-dependent process. Oncogene 23:1364–1376

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

We are grateful to Scott Kaufmann, M.D., Ph.D., and Eugenie Kleinerman, M.D., for helpful critical discussions. We also thank the M.D. Anderson Institutional Research Grant Program for support of studies examining adaphostin’s cytotoxicity.

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Authors and Affiliations

  1. Department of Pediatrics Research, Division of Pediatrics, University of Texas M.D. Anderson Cancer Center, 1515 Holcombe Blvd., 853, Houston, TX, 77030, USA

    Jason Long, Tejas Manchandia, Kechen Ban, Shan Gao, Claudia Miller & Joya Chandra

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  1. Jason Long
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  2. Tejas Manchandia
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Correspondence to Joya Chandra.

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Long, J., Manchandia, T., Ban, K. et al. Adaphostin cytoxicity in glioblastoma cells is ROS-dependent and is accompanied by upregulation of heme oxygenase-1. Cancer Chemother Pharmacol 59, 527–535 (2007). https://doi.org/10.1007/s00280-006-0295-5

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  • DOI: https://doi.org/10.1007/s00280-006-0295-5

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