Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Targeting MEK5 impairs nonhomologous end-joining repair and sensitizes prostate cancer to DNA damaging agents

Oncogene volume 39, pages 2467–2477 (2020)Cite this article

Subjects

Abstract

Radiotherapy is commonly used to treat a variety of solid human tumors, including localized prostate cancer. However, treatment failure often ensues due to tumor intrinsic or acquired radioresistance. Here we find that the MEK5/ERK5 signaling pathway is associated with resistance to genotoxic stress in aggressive prostate cancer cells. MEK5 knockdown by RNA interference sensitizes prostate cancer cells to ionizing radiation (IR) and etoposide treatment, as assessed by clonogenic survival and short-term proliferation assays. Mechanistically, MEK5 downregulation impairs phosphorylation of the catalytic subunit of DNA-PK at serine 2056 in response to IR or etoposide treatment. Although MEK5 knockdown does not influence the initial appearance of radiation- and etoposide-induced γH2AX and 53BP1 foci, it markedly delays their resolution, indicating a DNA repair defect. A cell-based assay shows that nonhomologous end joining (NHEJ) is compromised in cells with ablated MEK5 protein expression. Finally, MEK5 silencing combined with focal irradiation causes strong inhibition of tumor growth in mouse xenografts, compared with MEK5 depletion or radiation alone. These findings reveal a convergence between MEK5 signaling and DNA repair by NHEJ in conferring resistance to genotoxic stress in advanced prostate cancer and suggest targeting MEK5 as an effective therapeutic intervention in the management of this disease.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: MEK5 silencing sensitizes cells to radiation.
Fig. 2: MEK5 knockdown impairs DNA-PKcs phosphorylation in response to ionizing radiation.
Fig. 3: MEK5 knockdown impairs DNA-PKcs phosphorylation in response to etoposide and phleomycin.
Fig. 4: MEK5 knockdown delays resolution of irradiation-induced DSBs.
Fig. 5: MEK5 knockdown delays resolution of etoposide-induced DSBs.
Fig. 6: MEK5 depletion impairs non-homologous end joining.
Fig. 7

Similar content being viewed by others

References

  1. Bolla M, Van Tienhoven G, Warde P, Dubois JB, Mirimanoff RO, Storme G, et al. External irradiation with or without long-term androgen suppression for prostate cancer with high metastatic risk: 10-year results of an EORTC randomised study. Lancet Oncol. 2010;11:1066–73.

    Article  CAS  PubMed  Google Scholar 

  2. Palacios DA, Miyake M, Rosser CJ. Radiosensitization in prostate cancer: mechanisms and targets. BMC Urol. 2013;13:4.

    Article  PubMed  PubMed Central  Google Scholar 

  3. Ciccia A, Elledge SJ. The DNA damage response: making it safe to play with knives. Mol Cell. 2010;40:179–204.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. San Filippo J, Sung P, Klein H. Mechanism of eukaryotic homologous recombination. Annu Rev Biochem. 2008;77:229–57.

    Article  CAS  PubMed  Google Scholar 

  5. Lieber MR. The mechanism of human nonhomologous DNA end joining. J Biol Chem. 2008;283:1–5.

    Article  CAS  PubMed  Google Scholar 

  6. Jeggo PA. DNA breakage and repair. Adv Genet. 1998;38:185–218.

    Article  CAS  PubMed  Google Scholar 

  7. Davis AJ, Chen BP, Chen DJ. DNA-PK: a dynamic enzyme in a versatile DSB repair pathway. DNA Repair. 2014;17:21–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Chen BP, Chan DW, Kobayashi J, Burma S, Asaithamby A, Morotomi-Yano K, et al. Cell cycle dependence of DNA-dependent protein kinase phosphorylation in response to DNA double strand breaks. J Biol Chem. 2005;280:14709–15.

    Article  CAS  PubMed  Google Scholar 

  9. Zhou G, Bao ZQ, Dixon JE. Components of a new human protein kinase signal transduction pathway. J Biol Chem. 1995;270:12665–9.

    Article  CAS  PubMed  Google Scholar 

  10. Sun W, Kesavan K, Schaefer BC, Garrington TP, Ware M, Johnson NL, et al. MEKK2 associates with the adapter protein Lad/RIBP and regulates the MEK5-BMK1/ERK5 pathway. J Biol Chem. 2001;276:5093–100.

    Article  CAS  PubMed  Google Scholar 

  11. Chao TH, Hayashi M, Tapping RI, Kato Y, Lee JD. MEKK3 directly regulates MEK5 activity as part of the big mitogen-activated protein kinase 1 (BMK1) signaling pathway. J Biol Chem. 1999;274:36035–8.

    Article  CAS  PubMed  Google Scholar 

  12. Suzaki Y, Yoshizumi M, Kagami S, Koyama AH, Taketani Y, Houchi H, et al. Hydrogen peroxide stimulates c-Src-mediated big mitogen-activated protein kinase 1 (BMK1) and the MEF2C signaling pathway in PC12 cells: potential role in cell survival following oxidative insults. J Biol Chem. 2002;277:9614–21.

    Article  CAS  PubMed  Google Scholar 

  13. Arnoux V, Nassour M, L’Helgoualc’h A, Hipskind RA, Savagner P. Erk5 controls Slug expression and keratinocyte activation during wound healing. Mol Biol Cell. 2008;19:4738–49.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Zhou C, Nitschke AM, Xiong W, Zhang Q, Tang Y, Bloch M, et al. Proteomic analysis of tumor necrosis factor-alpha resistant human breast cancer cells reveals a MEK5/Erk5-mediated epithelial-mesenchymal transition phenotype. Breast Cancer Res. 2008;10:R105.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Schweppe RE, Cheung TH, Ahn NG. Global gene expression analysis of ERK5 and ERK1/2 signaling reveals a role for HIF-1 in ERK5-mediated responses. J Biol Chem. 2006;281:20993–1003.

    Article  CAS  PubMed  Google Scholar 

  16. English JM, Vanderbilt CA, Xu S, Marcus S, Cobb MH. Isolation of MEK5 and differential expression of alternatively spliced forms. J Biol Chem. 1995;270:28897–902.

    Article  CAS  PubMed  Google Scholar 

  17. Seyfried J, Wang X, Kharebava G, Tournier C. A novel mitogen-activated protein kinase docking site in the N terminus of MEK5alpha organizes the components of the extracellular signal-regulated kinase 5 signaling pathway. Mol Cell Biol. 2005;25:9820–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Cameron SJ, Abe J, Malik S, Che W, Yang J. Differential role of MEK5alpha and MEK5beta in BMK1/ERK5 activation. J Biol Chem. 2004;279:1506–12.

    Article  CAS  PubMed  Google Scholar 

  19. Tusa I, Gagliardi S, Tubita A, Pandolfi S, Urso C, Borgognoni L, et al. ERK5 is activated by oncogenic BRAF and promotes melanoma growth. Oncogene. 2018;37:2601–14.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Benito-Jardón L, Díaz-Martínez M, Arellano-Sánchez N, Vaquero-Morales P, Esparís-Ogando A, Teixidó J. Resistance to MAPK inhibitors in melanoma involves activation of the IGF1R-MEK5-Erk5 pathway. Cancer Res. 2019;79:2244–56.

    Article  PubMed  Google Scholar 

  21. Ghosh AK, Steele R, Ray RB. c-myc Promoter-binding protein 1 (MBP-1) regulates prostate cancer cell growth by inhibiting MAPK pathway. J Biol Chem. 2005;280:14325–30.

    Article  CAS  PubMed  Google Scholar 

  22. Regan CP, Li W, Boucher DM, Spatz S, Su MS, Kuida K. Erk5 null mice display multiple extraembryonic vascular and embryonic cardiovascular defects. Proc Natl Acad Sci USA. 2002;99:9248–53.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Yan L, Carr J, Ashby PR, Murry-Tait V, Thompson C, Arthur JS. Knockout of ERK5 causes multiple defects in placental and embryonic development. BMC Dev Biol. 2003;3:11.

    Article  PubMed  PubMed Central  Google Scholar 

  24. Loveridge CJ, Mui EJ, Patel R, Tan EH, Ahmad I, Welsh M, et al. Increased T-cell Infiltration elicited by Erk5 deletion in a Pten-deficient mouse model of prostate carcinogenesis. Cancer Res. 2017;77:3158–68.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Mehta PB, Jenkins BL, McCarthy L, Thilak L, Robson CN, Neal DE, et al. MEK5 overexpression is associated with metastatic prostate cancer, and stimulates proliferation, MMP-9 expression and invasion. Oncogene. 2003;22:1381–9.

    Article  CAS  PubMed  Google Scholar 

  26. McCracken SR, Ramsay A, Heer R, Mathers ME, Jenkins BL, Edwards J, et al. Aberrant expression of extracellular signal-regulated kinase 5 in human prostate cancer. Oncogene. 2008;27:2978–88.

    Article  CAS  PubMed  Google Scholar 

  27. Ramsay AK, McCracken SR, Soofi M, Fleming J, Yu AX, Ahmad I, et al. ERK5 signalling in prostate cancer promotes an invasive phenotype. Br J Cancer. 2011;104:664–72.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Adachi N, Suzuki H, Liizumi S, Koyama H. Hypersensitivity of nonhomologous DNA end-joining mutants to VP-16 and ICRF-193: implications for the repair of topoisomerase II-mediated DNA damage. J Biol Chem. 2003;278:35897–902.

    Article  CAS  PubMed  Google Scholar 

  29. Panier S, Durocher D. Push back to respond better: regulatory inhibition of the DNA double-strand break response. Nat Rev Mol Cell Biol. 2013;14:661–72.

    Article  CAS  PubMed  Google Scholar 

  30. Wang HC, Chou WC, Shieh SY, Shen CY. Ataxia telangiectasia mutated and checkpoint kinase 2 regulate BRCA1 to promote the fidelity of DNA end-joining. Cancer Res. 2006;66:1391–400.

    Article  CAS  PubMed  Google Scholar 

  31. Kim MR, Lee J, An YS, Jin YB, Park IC, Chung E, et al. TGFβ1 protects cells from γ-IR by enhancing the activity of the NHEJ repair pathway. Mol Cancer Res. 2015;13:319–29.

    Article  CAS  PubMed  Google Scholar 

  32. Wu CC, Chaudhary KR, Na YH, Welch D, Black PJ, Sonabend AM, et al. Quality assessment of stereotactic radiosurgery of a melanoma brain metastases model using a mouselike phantom and the small animal radiation research platform. Int J Radiat Oncol Biol Phys. 2017;99:191–201.

    Article  PubMed  PubMed Central  Google Scholar 

  33. Polkinghorn WR, Parker JS, Lee MX, Kass EM, Spratt DE, Iaquinta PJ, et al. Androgen receptor signaling regulates DNA repair in prostate cancers. Cancer Discov. 2013;3:1245–53.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Goodwin JF, Schiewer MJ, Dean JL, Schrecengost RS, de Leeuw R, Han S, et al. A hormone-DNA repair circuit governs the response to genotoxic insult. Cancer Discov. 2013;3:1254–71.

    Article  CAS  PubMed  Google Scholar 

  35. Shibata A, Conrad S, Birraux J, Geuting V, Barton O, Ismail A, et al. Factors determining DNA double-strand break repair pathway choice in G2 phase. EMBO J. 2011;30:1079–92.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Brenner JC, Ateeq B, Li Y, Yocum AK, Cao Q, Asangani IA, et al. Mechanistic rationale for inhibition of poly(ADP-ribose) polymerase in ETS gene fusion-positive prostate cancer. Cancer Cell. 2011;19:664–78.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Ni X, Zhang Y, Ribas J, Chowdhury WH, Castanares M, Zhang Z, et al. Prostate-targeted radiosensitization via aptamer-shRNA chimeras in human tumor xenografts. J Clin Investig. 2011;121:2383–90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Bouchaert P, Guerif S, Debiais C, Irani J. Fromont G DNA-PKcs expression predicts response to radiotherapy in prostate cancer. Int J Radiat Oncol Biol Phys. 2012;84:1179–85.

  39. Collis SJ, Swartz MJ, Nelson WG, DeWeese TL. Enhanced radiation and chemotherapy-mediated cell killing of human cancer cells by small inhibitory RNA silencing of DNA repair factors. Cancer Res. 2003;63:1550–4.

    CAS  PubMed  Google Scholar 

  40. Banáth JP, Macphail SH, Olive PL. Radiation sensitivity, H2AX phosphorylation, and kinetics of repair of DNA strand breaks in irradiated cervical cancer cell lines. Cancer Res. 2004;64:7144–9.

    Article  PubMed  Google Scholar 

  41. Olive PL, Banáth JP. Phosphorylation of histone H2AX as a measure of radiosensitivity. Int J Radiat Oncol Biol Phys. 2004;58:331–5.

    Article  CAS  PubMed  Google Scholar 

  42. Taneja N, Davis M, Choy JS, Beckett MA, Singh R, Kron SJ, et al. Histone H2AX phosphorylation as a predictor of radiosensitivity and target for radiotherapy. J Biol Chem. 2004;279:2273–80.

    Article  CAS  PubMed  Google Scholar 

  43. Stiff T, O’Driscoll M, Rief N, Iwabuchi K, Löbrich M, Jeggo PA. ATM and DNA-PK function redundantly to phosphorylate H2AX after exposure to ionizing radiation. Cancer Res. 2004;64:2390–6.

    Article  CAS  PubMed  Google Scholar 

  44. Nyati MK, Feng FY, Maheshwari D, Varambally S, Zielske SP, Ahsan A, et al. Ataxia telangiectasia mutated down-regulates phospho-extracellular signal-regulated kinase 1/2 via activation of MKP-1 in response to radiation. Cancer Res. 2006;66:11554–9.

    Article  CAS  PubMed  Google Scholar 

  45. Tang D, Wu D, Hirao A, Lahti JM, Liu L, Mazza B, et al. ERK activation mediates cell cycle arrest and apoptosis after DNA damage independently of p53. J Biol Chem. 2002;277:12710–7.

    Article  CAS  PubMed  Google Scholar 

  46. Golding SE, Rosenberg E, Neill S, Dent P, Povirk LF, Valerie K. Extracellular signal-related kinase positively regulates ataxia telangiectasia mutated, homologous recombination repair, and the DNA damage response. Cancer Res. 2007;67:1046–53.

    Article  CAS  PubMed  Google Scholar 

  47. Estrada-Bernal A, Chatterjee M, Haque SJ, Yang L, Morgan MA, Kotian S, et al. MEK inhibitor GSK1120212-mediated radiosensitization of pancreatic cancer cells involves inhibition of DNA double-strand break repair pathways. Cell Cycle. 2015;14:3713–24.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Wei F, Yan J, Tang D, Lin X, He L, Xie Y, et al. Inhibition of ERK activation enhances the repair of double-stranded breaks via non-homologous end joining by increasing DNA-PKcs activation. Biochim Biophys Acta. 2013;1833:90–100.

    Article  CAS  PubMed  Google Scholar 

  49. Jiang W, Jin G, Cai F, Chen X, Cao N, Zhang X, et al. Extracellular signal-regulated kinase 5 increases radioresistance of lung cancer cells by enhancing the DNA damage response. Exp Mol Med. 2019;51:19.

    Article  CAS  PubMed Central  Google Scholar 

  50. Pavan S, Meyer-Schaller N, Diepenbruck M, Kalathur RKR, Saxena M, Christofori G. A kinome-wide high-content siRNA screen identifies MEK5-ERK5 signaling as critical for breast cancer cell EMT and metastasis. Oncogene. 2018;37:4197–213.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This work is supported by the Department of Defense Prostate Cancer Research Program, W81XWH-15-1-0296 (CGB). This study also used the resources of the Herbert Irving Comprehensive Cancer Center Flow Cytometry, Radiation Research, and Confocal and Specialized Microscopy Shared Resources funded in part through Center Grant P30CA013696. We thank Theresa Swayne and Laura Munteanu from the Confocal and Specialized Microscopy Shared Resource of the Irving Cancer Research Center (ICRC) and Drs. Siu-Hong Ho, Wei Wang, and Caisheng Lu from the Flow Cytometry Shared Resource of the Herbert Irving Comprehensive Cancer Center at Columbia University Irving Medical Center for their help in performing the immunofluorescence and flow cytometry experiments.

Author information

Authors and Affiliations

  1. Center for Radiological Research, Columbia University Vagelos College of Physicians and Surgeons, Columbia University Irving Medical Center, New York, NY, USA

    Constantinos G. Broustas, Axel J. Duval, Kunal R. Chaudhary & Howard B. Lieberman

  2. Biomedical Informatics Shared Resource, Herbert Irving Comprehensive Cancer Center and Department of Biomedical Informatics, Columbia University Irving Medical Center, New York, NY, USA

    Richard A. Friedman

  3. Department of Pathology and Cell Biology, Columbia University Irving Medical Center, New York, NY, USA

    Renu K. Virk

  4. Department of Environmental Health Sciences, Mailman School of Public Health, Columbia University, New York, NY, USA

    Howard B. Lieberman

Authors
  1. Constantinos G. Broustas
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Axel J. Duval
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kunal R. Chaudhary
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Richard A. Friedman
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Renu K. Virk
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Howard B. Lieberman
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Constantinos G. Broustas.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Broustas, C.G., Duval, A.J., Chaudhary, K.R. et al. Targeting MEK5 impairs nonhomologous end-joining repair and sensitizes prostate cancer to DNA damaging agents. Oncogene 39, 2467–2477 (2020). https://doi.org/10.1038/s41388-020-1163-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41388-020-1163-1

This article is cited by

Search

Advanced search

Quick links