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Nuclear Localization of Robo is Associated with Better Survival in Bladder Cancer

  • Original Article
  • Published:
Pathology & Oncology Research

Abstract

The Slit-Robo pathway has shown to be altered in several malignant diseases. However, its role in bladder cancer is poorly understood. Therefore, we aimed to assess the tissue expression of Robo1 and Robo4 as well as their ligand Slit2 in different stages of bladder cancer to explore possible changes of Slit-Robo signalling during the progression of bladder cancer. Robo1, Robo4 and Slit2 gene expressions were analyzed in 92 frozen bladder cancer tissue samples by using reverse transcription quantitative real-time PCR. Immunohistochemical analyses were performed on 149 formalin-fixed and paraffin-embedded bladder cancer tissue samples. Results were correlated with the clinical and follow-up data by performing both univariable and multivariable analyses. Robo1 and Robo4 nuclear staining intensitiy was significantly higher in low stage and low grade bladder cancer. Elevated Robo1 nuclear staining was associated with better disease-specific survival (DSS) (p = 0.045). Similarly, stronger Robo4 nuclear staining tended to be associated with longer DSS (p = 0.061). We found higher Robo1 and Slit2 gene expression levels in advanced stages of bladder cancer (p = 0.007 and p < 0.001). High Slit2 gene expression was correlated with significantly shorter DSS (p < 0.005), while Robo1 and Robo4 gene expressions were not associated with patients’ prognosis. Our results demonstrate that the nuclear expression of Robo1 and Robo4 is associated with a favourable prognosis suggesting that its translocation into the nucleus represent a posttranslational regulation process which may exhibit an antitumor effect in bladder cancer.

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References

  1. Stein JP, Skinner DG (2006) Radical cystectomy for invasive bladder cancer: long-term results of a standard procedure. World J Urol 24(3):296–304. https://doi.org/10.1007/s00345-006-0061-7

    Article  PubMed  Google Scholar 

  2. Robertson AG, Kim J, Al-Ahmadie H, Bellmunt J, Guo G, Cherniack AD, Hinoue T, Laird PW, Hoadley KA, Akbani R, Castro MAA, Gibb EA, Kanchi RS, Gordenin DA, Shukla SA, Sanchez-Vega F, Hansel DE, Czerniak BA, Reuter VE, Su X, de Sa Carvalho B, Chagas VS, Mungall KL, Sadeghi S, Pedamallu CS, Lu Y, Klimczak LJ, Zhang J, Choo C, Ojesina AI, Bullman S, Leraas KM, Lichtenberg TM, Wu CJ, Schultz N, Getz G, Meyerson M, Mills GB, McConkey DJ, Weinstein JN, Kwiatkowski DJ, Lerner SP (2017) Comprehensive molecular characterization of muscle-invasive bladder Cancer. Cell 171(3):540–556.e525. https://doi.org/10.1016/j.cell.2017.09.007

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Wang B, Xiao Y, Ding BB, Zhang N, Yuan X, Gui L, Qian KX, Duan S, Chen Z, Rao Y, Geng JG (2003) Induction of tumor angiogenesis by Slit-Robo signaling and inhibition of cancer growth by blocking Robo activity. Cancer Cell 4(1):19–29

    Article  Google Scholar 

  4. Dickinson RE, Dallol A, Bieche I, Krex D, Morton D, Maher ER, Latif F (2004) Epigenetic inactivation of SLIT3 and SLIT1 genes in human cancers. Br J Cancer 91(12):2071–2078. https://doi.org/10.1038/sj.bjc.6602222

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Dickinson RE, Duncan WC (2010) The SLIT-ROBO pathway: a regulator of cell function with implications for the reproductive system. Reproduction (Cambridge, England) 139(4):697–704. https://doi.org/10.1530/rep-10-0017

    Article  CAS  Google Scholar 

  6. Kaur S, Castellone MD, Bedell VM, Konar M, Gutkind JS, Ramchandran R (2006) Robo4 signaling in endothelial cells implies attraction guidance mechanisms. J Biol Chem 281(16):11347–11356. https://doi.org/10.1074/jbc.M508853200

    Article  CAS  PubMed  Google Scholar 

  7. Sundaresan V, Heppell-Parton A, Coleman N, Miozzo M, Sozzi G, Ball R, Cary N, Hasleton P, Fowler W, Rabbitts P (1995) Somatic genetic changes in lung cancer and precancerous lesions. Ann Oncol 6(Suppl 1):27–31 discussion 31-22

  8. Biankin AV, Waddell N, Kassahn KS, Gingras MC, Muthuswamy LB, Johns AL, Miller DK, Wilson PJ, Patch AM, Wu J, Chang DK, Cowley MJ, Gardiner BB, Song S, Harliwong I, Idrisoglu S, Nourse C, Nourbakhsh E, Manning S, Wani S, Gongora M, Pajic M, Scarlett CJ, Gill AJ, Pinho AV, Rooman I, Anderson M, Holmes O, Leonard C, Taylor D, Wood S, Xu Q, Nones K, Fink JL, Christ A, Bruxner T, Cloonan N, Kolle G, Newell F, Pinese M, Mead RS, Humphris JL, Kaplan W, Jones MD, Colvin EK, Nagrial AM, Humphrey ES, Chou A, Chin VT, Chantrill LA, Mawson A, Samra JS, Kench JG, Lovell JA, Daly RJ, Merrett ND, Toon C, Epari K, Nguyen NQ, Barbour A, Zeps N, Kakkar N, Zhao F, Wu YQ, Wang M, Muzny DM, Fisher WE, Brunicardi FC, Hodges SE, Reid JG, Drummond J, Chang K, Han Y, Lewis LR, Dinh H, Buhay CJ, Beck T, Timms L, Sam M, Begley K, Brown A, Pai D, Panchal A, Buchner N, De Borja R, Denroche RE, Yung CK, Serra S, Onetto N, Mukhopadhyay D, Tsao MS, Shaw PA, Petersen GM, Gallinger S, Hruban RH, Maitra A, Iacobuzio-Donahue CA, Schulick RD, Wolfgang CL, Morgan RA, Lawlor RT, Capelli P, Corbo V, Scardoni M, Tortora G, Tempero MA, Mann KM, Jenkins NA, Perez-Mancera PA, Adams DJ, Largaespada DA, Wessels LF, Rust AG, Stein LD, Tuveson DA, Copeland NG, Musgrove EA, Scarpa A, Eshleman JR, Hudson TJ, Sutherland RL, Wheeler DA, Pearson JV, McPherson JD, Gibbs RA, Grimmond SM (2012) Pancreatic cancer genomes reveal aberrations in axon guidance pathway genes. Nature 491(7424):399–405. https://doi.org/10.1038/nature11547

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Gara RK, Kumari S, Ganju A, Yallapu MM, Jaggi M, Chauhan SC (2015) Slit/Robo pathway: a promising therapeutic target for cancer. Drug Discov Today 20(1):156–164. https://doi.org/10.1016/j.drudis.2014.09.008

    Article  CAS  PubMed  Google Scholar 

  10. Prasad A, Fernandis AZ, Rao Y, Ganju RK (2004) Slit protein-mediated inhibition of CXCR4-induced chemotactic and chemoinvasive signaling pathways in breast cancer cells. J Biol Chem 279(10):9115–9124. https://doi.org/10.1074/jbc.M308083200

    Article  CAS  PubMed  Google Scholar 

  11. Prasad A, Paruchuri V, Preet A, Latif F, Ganju RK (2008) Slit-2 induces a tumor-suppressive effect by regulating beta-catenin in breast cancer cells. J Biol Chem 283(39):26624–26633. https://doi.org/10.1074/jbc.M800679200

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods (San Diego, Calif) 25(4):402–408. https://doi.org/10.1006/meth.2001.1262

    Article  CAS  Google Scholar 

  13. Huang T, Kang W, Cheng AS, Yu J, To KF (2015) The emerging role of Slit-Robo pathway in gastric and other gastro intestinal cancers. BMC Cancer 15:950. https://doi.org/10.1186/s12885-015-1984-4

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Legg JA, Herbert JM, Clissold P, Bicknell R (2008) Slits and roundabouts in cancer, tumour angiogenesis and endothelial cell migration. Angiogenesis 11(1):13–21. https://doi.org/10.1007/s10456-008-9100-x

    Article  PubMed  Google Scholar 

  15. Zhou WJ, Geng ZH, Chi S, Zhang W, Niu XF, Lan SJ, Ma L, Yang X, Wang LJ, Ding YQ, Geng JG (2011) Slit-Robo signaling induces malignant transformation through Hakai-mediated E-cadherin degradation during colorectal epithelial cell carcinogenesis. Cell Res 21(4):609–626. https://doi.org/10.1038/cr.2011.17

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Stella MC, Trusolino L, Comoglio PM (2009) The Slit/Robo system suppresses hepatocyte growth factor-dependent invasion and morphogenesis. Mol Biol Cell 20(2):642–657. https://doi.org/10.1091/mbc.E08-03-0321

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Marlow R, Strickland P, Lee JS, Wu X, Pebenito M, Binnewies M, Le EK, Moran A, Macias H, Cardiff RD, Sukumar S, Hinck L (2008) SLITs suppress tumor growth in vivo by silencing Sdf1/Cxcr4 within breast epithelium. Cancer Res 68(19):7819–7827. https://doi.org/10.1158/0008-5472.can-08-1357

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Schmid BC, Rezniczek GA, Fabjani G, Yoneda T, Leodolter S, Zeillinger R (2007) The neuronal guidance cue Slit2 induces targeted migration and may play a role in brain metastasis of breast cancer cells. Breast Cancer Res Treat 106(3):333–342. https://doi.org/10.1007/s10549-007-9504-0

    Article  PubMed  Google Scholar 

  19. Li Y, Cheng H, Xu W, Tian X, Li X, Zhu C (2015) Expression of Robo protein in bladder cancer tissues and its effect on the growth of cancer cells by blocking Robo protein. Int J Clin Exp Pathol 8(9):9932–9940

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Fujiwara M, Ghazizadeh M, Kawanami O (2006) Potential role of the Slit/Robo signal pathway in angiogenesis. Vasc Med (London, England) 11(2):115–121

    Google Scholar 

  21. Seki M, Watanabe A, Enomoto S, Kawamura T, Ito H, Kodama T, Hamakubo T, Aburatani H (2010) Human ROBO1 is cleaved by metalloproteinases and gamma-secretase and migrates to the nucleus in cancer cells. FEBS Lett 584(13):2909–2915. https://doi.org/10.1016/j.febslet.2010.05.009

    Article  CAS  PubMed  Google Scholar 

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Funding

This work was supported by the National Research, Development and Innovation Office – NKFIH / PD 115616. Tibor Szarvas was supported by János Bolyai Research Scholarship of the Hungarian Academy of Sciences.

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Authors and Affiliations

  1. Department of Urology, Faculty of Medicine, University Duisburg-Essen, Hufelandstr 55, 45147, Essen, Germany

    Ulrich Krafft, Markus Becker, Christian Niedworok, Christopher Darr, Boris Hadaschik & Tibor Szarvas

  2. Institute of Pathology, Faculty of Medicine, University Duisburg-Essen, Essen, Germany

    Henning Reis & Marc Ingenwerth

  3. 1st Institute of Pathology and Experimental Cancer Research, Semmelweis University, Budapest, Hungary

    Ilona Kovalszky

  4. Department of Urology, Semmelweis University, Üllöi út 78/b, Budapest, 1082, Hungary

    Péter Nyirády & Tibor Szarvas

Authors
  1. Ulrich Krafft
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  2. Henning Reis
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  3. Marc Ingenwerth
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  4. Ilona Kovalszky
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  5. Markus Becker
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  6. Christian Niedworok
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  7. Christopher Darr
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  8. Péter Nyirády
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  9. Boris Hadaschik
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  10. Tibor Szarvas
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Correspondence to Tibor Szarvas.

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All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

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Krafft, U., Reis, H., Ingenwerth, M. et al. Nuclear Localization of Robo is Associated with Better Survival in Bladder Cancer. Pathol. Oncol. Res. 26, 253–261 (2020). https://doi.org/10.1007/s12253-018-0447-z

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  • DOI: https://doi.org/10.1007/s12253-018-0447-z

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