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Mutant TP53 enhances the resistance of glioblastoma cells to temozolomide by up-regulating O6-methylguanine DNA-methyltransferase

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Abstract

The “gain of function” of mutant TP53 is an important determinant in human tumor development and progression. This study aimed to investigate the possible mechanism of mutant TP53 inducing temozolomide resistance in glioblastoma cells. Three established human glioma cell lines, T98G, U87, and U138, were chemoresistant cells. The mRNA of cells was sequenced to confirm the status of TP53. Synthetic small interfering RNA (siRNA) was used to knock down TP53 in cells. TP53 mRNA was detected “silenced” by reverse transcriptase-polymerase chain reaction (RT-PCR) in five consecutive days. Viable cell survival was measured when these cells were exposed to temozolomide or semustine in step-up concentrations. The expression of O6-methylguanine DNA-methyltransferase (MGMT) at mRNA level was also determined. T98G, U87, and U138 cells were resistant to temozolomide. T98G and U138 cells expressed mutant-type TP53 with positive MGMT, while U87 cell expressed wild-type TP53 with negative MGMT. TP53-siRNA knocked down TP53 effectively (P = 0.021) in five consecutive days. Knockdown of mutant TP53 in T98G and U138 cells led to a fivefold increase in chemosensitivity to temozolomide, but not semustine. Knockdown of wild TP53 in U87 cell did not affect the chemoresistance. In addition, mutant TP53 knockdown induced a dramatic decrease of MGMT expression (P = 0.0000034). TP53 mutation decreases the chemosensitivity of malignant gliomas to temozolomide. This “gain of function” in drug resistance may be obtained by increasing MGMT expression.

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References

  1. Stupp R, Hegi ME, Mason WP, van den Bent MJ, Taphoorn MJ, Janzer RC, Ludwin SK, Allgeier A, Fisher B, Belanger K, Hau P, Brandes AA, Gijtenbeek J, Marosi C, Vecht CJ, Mokhtari K, Wesseling P, Villa S, Eisenhauer E, Gorlia T, Weller M, Lacombe D, Cairncross JG, Mirimanoff RO et al (2009) European Organisation for Research and Treatment of Cancer Brain Tumour and Radiation Oncology Groups, National Cancer Institute of Canada Clinical Trials Group. Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial. Lancet Oncol 10:459–466

    Article  PubMed  CAS  Google Scholar 

  2. Mirimanoff RO, Gorlia T, Mason W, Van den Bent MJ, Kortmann RD, Fisher B, Reni M, Brandes AA, Curschmann J, Villa S, Cairncross G, Allgeier A, Lacombe D, Stupp R (2006) Radiotherapy and temozolomide for newly diagnosed glioblastoma: recursive partitioning analysis of the EORTC 26981/22981-NCIC CE3 phase III randomized trial. J Clin Oncol 24:2563–2569

    Article  PubMed  CAS  Google Scholar 

  3. Hegi ME, Diserens AC, Gorlia T, Hamou MF, de Tribolet N, Weller M, Kros JM, Hainfellner JA, Mason W, Mariani L, Bromberg JE, Hau P, Mirimanoff RO, Cairncross JG, Janzer RC, Stupp R (2005) MGMT gene silencing and benefit from temozolomide in glioblastoma. N Engl J Med 352:997–1003

    Article  PubMed  CAS  Google Scholar 

  4. Louis DN, von Deimling A, Chung RY, Rubio MP, Whaley JM, Eibl RH, Ohgaki H, Wiestler OD, Thor AD, Seizinger BR (1993) Comparative study of p53 gene and protein alterations in human astrocytic tumors. J Neuropathol Exp Neurol 52:31–38

    Article  PubMed  CAS  Google Scholar 

  5. Deppert W (2007) Mutant TP53: from guardian to fallen angel? Oncogene 26:2142–2144

    Article  PubMed  CAS  Google Scholar 

  6. Sampath J, Sun D, Kidd VJ, Grenet J, Gandhi A, Shapiro LH, Wang Q, Zambetti GP, Schuetz JD (2001) Mutant TP53 cooperates with ETS and selectively up-regulates human MDR1 not MRP1. J Biol Chem 276:39359–39367

    Article  PubMed  CAS  Google Scholar 

  7. Bossi G, Lapi E, Strano S, Rinaldo C, Blandino G, Sacchi A (2006) Mutant TP53 gain of function: reduction of tumor malignancy of human cancer cell lines through abrogation of mutant TP53 expression. Oncogene 25:304–309

    PubMed  CAS  Google Scholar 

  8. Tsang WP, Ho FY, Fung KP, Kong SK, Kwok TT (2005) TP53-175H mutant gains new function in regulation of doxorubicin-induced apoptosis. Int J Cancer 114:331–336

    Article  PubMed  CAS  Google Scholar 

  9. Zalcenstein A, Stambolsky P, Weisz L, Müller M, Wallach D, Goncharov TM, Krammer PH, Rotter V, Oren M (2003) Mutant TP53 gain of function: repression of CD95 (Fas/APO-1) gene expression by tumor associated TP53 mutants. Oncogene 22:5667–5676

    Article  PubMed  CAS  Google Scholar 

  10. Blandino G, Levine AJ, Oren M (1999) Mutant TP53 gain of function: differential effects of different TP53 mutants on resistance of cultured cells to chemotherapy. Oncogene 18:477–485

    Article  PubMed  CAS  Google Scholar 

  11. Natsume A, Wakabayashi T, Ishii D, Maruta H, Fujii M, Shimato S, Ito M, Yoshida J (2008) A combination of IFN-beta and temozolomide in human glioma xenograft models: implication of TP53-mediated MGMT downregulation. Cancer Chemother Pharmacol 61:653–659

    Article  PubMed  CAS  Google Scholar 

  12. Hermisson M, Klumpp A, Wick W, Wischhusen J, Nagel G, Roos W, Kaina B, Weller M (2006) O6-Methylguanine DNA methyltransferase and TP53 status predict temozolomide sensitivity in human malignant glioma cells. J Neurochem 96:766–776

    Article  PubMed  CAS  Google Scholar 

  13. Halevy O, Michalovitz D, Oren M (1990) Different tumor-derived TP53 mutant exhibit distinct biological activities. Science 250:113–116

    Article  PubMed  CAS  Google Scholar 

  14. Selivanova G, Wiman KG (2007) Reactivation of mutant TP53, molecular mechanisms and therapeutic potential. Oncogene 26:2243–2254

    Article  PubMed  CAS  Google Scholar 

  15. Soussi T (2007) TP53 alterations in human cancer: more questions than answers. Oncogene 26:2145–2156

    Article  PubMed  CAS  Google Scholar 

  16. Osanai T, Takagi Y, Toriya Y, Nakagawa T, Aruga T, Iida S, Uetake H, Sugihara K (2005) Inverse correlation between the expression of O6-methylguanine-DNA methyltransferase (MGMT) and TP53 in breast cancer. Jpn J Clin Oncol 35:121–125

    Article  PubMed  Google Scholar 

  17. Grombacher T, Eichhorn U, Kaina B (1998) TP53 is involved in regulation of the DNA repair gene O6-methylguanine-DNA methyltransferase (MGMT) by DNA damaging agents. Oncogene 17:845–851

    Article  PubMed  CAS  Google Scholar 

  18. Blough MD, Zlatescu MC, Cairncross JG (2007) O6-Methylguanine-DNA methyltransferase regulation by TP53 in astrocytic cells. Cancer Res 67:580–584

    Article  PubMed  CAS  Google Scholar 

  19. Petitjean A, Mathe E, Kato S, Ishioka C, Tavtigian SV, Hainaut P, Olivier M (2007) Impact of mutant TP53 functional properties on TP53 mutation patterns and tumor phenotype: lessons from recent developments in the IARC TP53 database. Hum Mutat 28:622–629

    Article  PubMed  CAS  Google Scholar 

  20. Xu GW, Mymryk JS, Cairncross JG (2005) Pharmaceutical mediated inactivation of TP53 sensitizes U87MG glioma cells to BCNU and temozolomide. Int J Cancer 116:187–192

    Article  PubMed  CAS  Google Scholar 

  21. Yuan Q, Matsumoto K, Nakabeppu Y, Iwaki T (2003) A comparative immunohistochemistry of O6-methylguanine-DNA methyltransferase and TP53 in diffusely infiltrating astrocytomas. Neuropathol 23:203–209

    Article  Google Scholar 

  22. Foroutan B, Ali Ruf A, Jerwood D, Anderson D et al (2007) In vitro studies of DNA damage and its repair in cells from NHL patients with different TP53 mutant protein status, resistant (TP53(+)) and sensitive (TP53(−)) to cancer chemotherapy. J Pharmacol Toxicol Methods 55:58–64

    Article  PubMed  CAS  Google Scholar 

  23. Groenendijk FH, Taal W, Dubbink HJ, Haarloo CR, Kouwenhoven MC, van den Bent MJ, Kros JM, Dinjens WN (2011) MGMT promoter hypermethylation is a frequent, early, and consistent event in astrocytoma progression, and not correlated with TP53 mutation. J Neurooncol 101:405–417

    Article  PubMed  CAS  Google Scholar 

  24. Weisz L, Damalas A, Liontos M, Karakaidos P, Fontemaggi G, Maor-Aloni R, Kalis M, Levrero M, Strano S, Gorgoulis VG, Rotter V, Blandino G, Oren M (2007) Mutant TP53 enhances nuclear factor kappaB activation by tumor necrosis factor alpha in cancer cells. Cancer Res 67:2396–2401

    Article  PubMed  CAS  Google Scholar 

  25. Petitjean A, Achatz MIW, Borresen-Dale AL, Hainaut P, Olivier M (2007) TP53 mutations in human cancers: functional selection and impact on cancer prognosis and outcomes. Oncogene 26:2157–2165

    Article  PubMed  CAS  Google Scholar 

  26. Wong RP, Tsang WP, Chau PY, Co NN, Tsang TY, Kwok TT (2007) TP53-R273H gains new function in induction of drug resistance through down-regulation of procaspase-3. Mol Cancer Ther 6:1054–1061

    Article  PubMed  CAS  Google Scholar 

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The authors declare that they have no conflict of interests.

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Authors and Affiliations

  1. Department of Neurosurgery, West China Hospital, Sichuan University, 37 Guo Xuexiang, Chengdu, 610041, Sichuan, China

    Xiang Wang, Yan-hui Liu, Chao You & Qing Mao

  2. Department of Radiology, Sichuan Cancer Hospital, Chengdu, China

    Jin-xiu Chen

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  1. Xiang Wang
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  2. Jin-xiu Chen
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  3. Yan-hui Liu
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  4. Chao You
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  5. Qing Mao
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Correspondence to Qing Mao.

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Wang, X., Chen, Jx., Liu, Yh. et al. Mutant TP53 enhances the resistance of glioblastoma cells to temozolomide by up-regulating O6-methylguanine DNA-methyltransferase. Neurol Sci 34, 1421–1428 (2013). https://doi.org/10.1007/s10072-012-1257-9

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  • DOI: https://doi.org/10.1007/s10072-012-1257-9

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