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HJURP knockdown disrupts clonogenic capacity and increases radiation-induced cell death of glioblastoma cells

Cancer Gene Therapy volume 27, pages 319–329 (2020)Cite this article

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Abstract

The Holliday Junction-Recognition Protein (HJURP) was reported as overexpressed in several cancers and also strongly correlated with poor prognosis of patients, especially in glioblastoma (GBM), the most common and deadly type of primary brain tumor. HJURP is responsible for loading the histone H3 variant—the Centromeric Protein A (CENP-A)—at the centromeres in a cell cycle-regulated manner, being required for proper chromosome segregation. Here we investigated HJURP association with survival and radioresistance of different GBM cell lines. HJURP knockdown compromised the clonogenic capacity and severely impaired survival of five distinct GBM cells, while nontumor astrocytes were not affected. U251MG cells showed a robust cell cycle arrest in G2/M phases followed by a drastic increment in cell death after HJURP silencing, while U138MG and U343MG cell lines presented augmented senescence with a comparable increase in cell death. Importantly, we verified that the impact on cell cycle dynamics and clonogenic survival were associated with loss CENP-A at the centromeres. Moreover, radiation resistance was also impacted by HJURP modulation in several GBM cell lines. U87MG, T98G, U138MG, and U343MG cells were all sensitized to ionizing radiation after HJURP reduction. These data reinforce the requirement of HJURP for proliferative capacity and radioresistance of tumor cells, underlining its potential as a promising therapeutic target for GBM.

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References

  1. Dunleavy EM, Roche D, Tagami H, Lacoste N, Ray-Gallet D, Nakamura Y, et al. HJURP Is a cell-cycle-dependent maintenance and deposition factor of CENP-A at centromeres. Cell. 2009;137:485–97.

    Article  CAS  Google Scholar 

  2. Foltz DR, Jansen LE, Bailey AO, Yates JR 3rd, Bassett EA, Wood S, et al. centromere-specific assembly of CENP-A nucleosomes is mediated by HJURP. Cell. 2009;137:472–84.

    Article  CAS  Google Scholar 

  3. Müller S, Montes de Oca R, Lacoste N, Dingli F, Loew D, Almouzni G. Phosphorylation and DNA binding of HJURP determine its centromeric recruitment and function in CenH3CENP-A loading. Cell Rep. 2014;8:190–203.

    Article  Google Scholar 

  4. Sekulic N, Black BE. Molecular underpinnings of centromere identity and maintenance. Trends Biochem Sci. 2012;37:220–29.

    Article  CAS  Google Scholar 

  5. Kato T, Sato N, Hayama S, Yamabuki T, Ito T, Miyamoto M, et al. Activation of holliday junction-recognizing protein involved in the chromosomal stability and immortality of cancer cells. Cancer Res. 2007;67:8544–53.

    Article  CAS  Google Scholar 

  6. Maehara K, Takahashi K, Saitoh S. CENP-A reduction induces a p53-Dependent cellular senescence response to protect cells from executing defective mitoses. Mol Cell Biol. 2010;30:2090–104.

    Article  CAS  Google Scholar 

  7. Heo JI, Cho JH, Kim JR. HJURP regulates cellular senescence in human fibroblasts and endothelial cells via a p53-dependent pathway. J Gerontol Ser A Biol Sci Med Sci. 2013;68:1–12.

    Article  Google Scholar 

  8. Hu Z, Huang G, Sadanandam A, Gu S, Lenburg ME, Pai M, et al. The expression level of HJURP has an independent prognostic impact and predicts the sensitivity to radiotherapy in breast cancer. Breast Cancer Res. 2010;12:R18.

    Article  Google Scholar 

  9. Hu B, Wang Q, Wang Y, Chen J, Li P, Han M. Holliday junction – recognizing protein promotes cell proliferation and correlates with unfavorable clinical outcome of hepatocellular carcinoma. Onco Targets Ther. 2017;10:2601–7.

    Article  Google Scholar 

  10. Li L, Li X, Meng Q, Khan AQ, Chen X. Increased expression of holliday junction- recognizing protein (HJURP) as an independent prognostic biomarker in advanced-stage serous ovarian carcinoma. Med Sci Monit. 2018;24:3050–5.

    Article  CAS  Google Scholar 

  11. Valente V, Serafim RB, de Oliveira LC, Adorni FS, Torrieri R, Tirapelli DP, et al. Modulation of HJURP (Holliday Junction-Recognizing Protein) levels is correlated with glioblastoma cells survival. PLoS ONE. 2013;8:e62200.

    Article  CAS  Google Scholar 

  12. Adamson C, Kanu OO, Mehta AI, Di C, Lin N, Mattox AK, et al. Glioblastoma multiforme: a review of where we have been and where we are going. Expert Opin Invest Drugs. 2009;18:1061–83.

    Article  CAS  Google Scholar 

  13. Jansen M, Yip S, Louis DN. Molecular pathology in adult gliomas: diagnostic, prognostic, and predictive markers. Lancet Neurol. 2010;9:717–26.

    Article  CAS  Google Scholar 

  14. Brandes AA, Ermani M, Basso U, Amistà P, Berti F, Scienza R, et al. Temozolomide as a second-line systemic regimen in recurrent high-grade glioma: a phase II study. Ann Oncol. 2001;12:255–57.

    Article  CAS  Google Scholar 

  15. Martinez R, Rohde V, Schackert G. Different molecular patterns in glioblastoma multiforme subtypes upon recurrence. J Neurooncol. 2010;96:321–29.

    Article  CAS  Google Scholar 

  16. Parsons DW, Jones S, Zhang X, Lin JC, Leary RJ, Angenendt P, et al. An integrated genomic analysis of human glioblastoma multiforme. Science. 2008;321:1807–12.

    Article  CAS  Google Scholar 

  17. Ishii N, Maier D, Merlo A, Tada M, Sawamura Y, Diserens AC, et al. Frequent co-alterations of TP53, p16/CDKN2A, p14ARF, PTEN tumor suppressor genes in human glioma cell lines. Brain Pathol. 1999;479:469–79.

    Article  Google Scholar 

  18. Barnhart MC, Kuich PH, Stellfox ME, Ward JA, Bassett EA, Black BE, et al. HJURP Is a CENP-A chromatin assembly factor sufficient to form a functional de novo kinetochore. J Cell Biol. 2011;194:229–43.

    Article  CAS  Google Scholar 

  19. Barnhart-Dailey MC, Trivedi P, Stukenberg PT, Foltz DR. HJURP interaction with the condensin II complex during G1 promotes CENP-A deposition. Mol Biol Cell. 2017;28:54–64.

    Article  CAS  Google Scholar 

  20. Mishra PK, Au WC, Choy JS, Kuich PH, Baker RE, Foltz DR, et al. Misregulation of Scm3p/HJURP causes chromosome instability in saccharomyces cerevisiae and human cells. PLoS Genet. 2011;7:e1002303.

    Article  CAS  Google Scholar 

  21. Perpelescu M, Hori T, Toyoda A, Misu S, Monma N, Ikeo K, et al. HJURP is involved in the expansion of centromeric chromatin. Mol Biol Cell. 2015;26:2742–54.

    Article  CAS  Google Scholar 

  22. Adams RR, Carmena M, Earnshaw WC. Chromosomal passengers and the (Aurora) ABCs of mitosis. Trends Cell Biol. 2001;11:49–54.

    Article  CAS  Google Scholar 

  23. Nigg EA. Mitotic kinases as regulators of cell division and its checkpoints. Nat Rev Mol Cell Biol. 2001;2:21–32.

    Article  CAS  Google Scholar 

  24. Keen N, Taylor S. Aurora-kinase inhibitors as anticancer agents. Nat Rev Cancer. 2004;4:927–36.

    Article  CAS  Google Scholar 

  25. Martens-de Kemp SR, Nagel R, Stigter-van Walsum M, van der Meulen IH, van Beusechem VW, Braakhuis BJ, et al. Functional genetic screens identify genes essential for tumor cell survival in head and neck and lung cancer. Clin Cancer Res. 2013;19:1994–2003.

    Article  CAS  Google Scholar 

  26. Chen C, Iavarone P, Fick A, Edwards J, Prados M, Israel MA. Constitutional p53 mutations associated with brain tumors in young adults. Cancer Genet Cytogenet. 1995;82:106–15.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors acknowledge Silvia Regina Andrade Nascimento for technical assistance, Fabiana Rossetto de Morais for flow cytometry analyses and Douglas Elias Santos for graphic art support in the illustration production.

Funding statement

VV was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo—FAPESP (grant #2013/13465–1; grant #2018/05018–9), Programa de Apoio ao Desenvolvimento Científico—PADC (grant no. 256/2017) from the Faculty of Pharmaceutical Sciences of Araraquara and by the Center for Cell-Based Therapy (CEPID/FAPESP; grant #2013/08135–2). RBS and LFMDC received fellowships from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—CAPES. CC received fellowships from the Conselho Nacional de Desenvolvimento Científico e Tecnológico—CNPq.

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Authors and Affiliations

  1. Department of Cellular and Molecular Biology, Ribeirão Preto Medical School, University of São Paulo (USP), Avenida Bandeirantes, 3900, Ribeirão Preto, 14049-900, Brazil

    Rodolfo B. Serafim, Cibele Cardoso, Luis F. M. Di Cristofaro, Enilza M. Espreafico, Maria L. Paçó-Larson & Valeria Valente

  2. São Paulo State University (UNESP), School of Pharmaceutical Sciences,, Rodovia Araraquara - Jaú, Km 01 - s/n, Campos Ville, Araraquara, SP, 14800-903, Brazil

    Christiane Pienna Soares & Valeria Valente

  3. Department of Genetics, Ribeirão Preto Medical School, University of São Paulo (USP), Avenida Bandeirantes, 3900, Ribeirão Preto, 14049-900, Brazil

    Wilson Araújo Silva Jr

  4. Center for Cell-Based Therapy-CEPID/FAPESP, Rua Tenente Catão Roxo, 2501, Ribeirão Preto, 14051-140, Brazil

    Wilson Araújo Silva Jr & Valeria Valente

  5. Center for Medical Genomics, HCFMRP/USP, and Center for Integrative System Biology - CISBi NAP/USP, University of São Paulo, Ribeirão Preto, São Paulo, Brazil

    Wilson Araújo Silva Jr

  6. Department of Radiation Oncology, Dana-Farber Cancer Institute, Boston, MA, 02215, USA

    Brendan D. Price

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  1. Rodolfo B. Serafim
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  2. Cibele Cardoso
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  9. Valeria Valente
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Contributions

Conceptualization: VV and RBS; Investigation: RBS, CC, and LFMDC; Funding acquisition: VV, CPS, WASJr, EME, and MLPL; Data analysis and writing: RBS., BDP and VV.

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Correspondence to Valeria Valente.

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Serafim, R.B., Cardoso, C., Di Cristofaro, L.F.M. et al. HJURP knockdown disrupts clonogenic capacity and increases radiation-induced cell death of glioblastoma cells. Cancer Gene Ther 27, 319–329 (2020). https://doi.org/10.1038/s41417-019-0103-0

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