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Loss of NF2/Merlin expression in advanced sporadic colorectal cancer

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Abstract

Purpose

NF2/Merlin was first identified through its association with neurofibromatosis type 2 (NF2). However, accumulating evidence suggests a more general involvement in tumorigenesis and, in particular, a broader role in tumor suppression. The aim of this study was to examine NF2/Merlin involvement in sporadic colorectal cancer.

Methods

This study is the first to examine the role of NF2/Merlin in sporadic colorectal cancer through LOH analysis at the NF2 locus and mRNA expression analysis via quantitative RT-PCR of total NF2, NF2 isoform I and II. In addition, Merlin protein expression was assessed by immunohistochemistry and Western blotting.

Results

NF2 LOH was detected in 20.0 % of heterozygous cases and was found to be more frequent in tumors larger than 5 cm in diameter (p = 0.041) and in tumors with a less differentiated phenotype (p = 0.027). No differences were observed in total NF2 and NF2 isoform I/isoform II mRNA expression between the tumors and their corresponding normal mucous tissues. NF2 isoform II was the most predominant isoform in all samples analyzed. mRNA expression levels of total NF2 and isoforms I and II were significantly lower in poorly differentiated tumors (p = 0.033, p = 0.036 and p = 0.044, respectively). Weak Merlin immunostaining was more frequent in poorly differentiated tumors (p = 0.034) and tumors classified as Dukes’ C (p = 0.023). A distinct pattern of Merin phosphorylation was observed in tumors compared to normal mucous tissues.

Conclusion

Our data indicate that NF2/Merlin may serve as a potential target in the management of colorectal cancer.

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References

  1. A. Jemal, F. Bray, M.M. Center, J. Ferlay, E. Ward, D. Forman, Global cancer statistics. CA Cancer J. Clin. 61, 69–90 (2011)

    Article  PubMed  Google Scholar 

  2. E.R. Fearon, Molecular genetics of colorectal cancer. Annu. Rev. Pathol. 6, 479–507 (2011)

    Article  CAS  PubMed  Google Scholar 

  3. J.A. Trofatter, M.M. MacCollin, J.L. Rutter, J.R. Murrell, M.P. Duyao, D.M. Parry, R. Eldridge, N. Kley, A.G. Menon, K. Pulaski, A novel moesin-, ezrin-, radixin-like gene is a candidate for the neurofibromatosis 2 tumor suppressor. Cell 72, 791–800 (1993)

    Article  CAS  PubMed  Google Scholar 

  4. T. Okada, L. You, F.G. Giancotti, Shedding light on Merlin’s wizardry. Trends Cell Biol. 17, 222–229 (2007)

    Article  CAS  PubMed  Google Scholar 

  5. Z.S. Morris, A.I. McClatchey, Aberrant epithelial morphology and persistent epidermal growth factor receptor signaling signaling in a mouse model of renal carcinoma. Proc. Natl. Acad. Sci. 106, 9767–9772 (2009)

    Article  CAS  PubMed  Google Scholar 

  6. H. Yu, Y. Zhang, L. Ye, W.G. Jiang, The FERM family proteins in cancer invasion and metastasis. Front. Biosci. 16, 1536–1550 (2011)

    Article  CAS  Google Scholar 

  7. I. Stamenkovic, Q. Yu, Merlin, a magic linker between the extracellular cues and intracelular signaling pathways that regulate cell motility, proliferation and survival. Curr. Protein Pept. Sci. 11, 471–484 (2010)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. A.B. Bianchi, T. Hara, V. Ramesh, J. Gao, A.J. Klein-Szanto, F. Morin, A.G. Menon, J.A. Trofatter, J.F. Gusella, B.R. Seizinger, N. Kley, Mutations in transcript isoforms of the neurofibromatosis 2 gene in multiple human tumour types. Nat. Genet. 6, 185–192 (1994)

    Article  CAS  PubMed  Google Scholar 

  9. L. Sherman, H.M. Xu, R.T. Geist, S. Saporito-Irwin, N. Howells, H. Ponta, P. Herrlich, D.H. Gutmann, Interdomain binding mediates tumor growth suppression by the NF2 gene product. Oncogene 15, 2505–2509 (1997)

    Article  CAS  PubMed  Google Scholar 

  10. D.H. Gutmann, C.A. Haipek, L.K. Hoang, Neurofibromatosis 2 tumor suppressor protein, merlin, forms two functionally important intramolecular associations. J. Neurosci. Res. 58, 706–716 (1999)

    Article  CAS  PubMed  Google Scholar 

  11. G.H. Xiao, A. Beeser, J. Chernoff, J.R. Testa, p21-activated kinase links Rac/Cdc42 signaling to merlin. J. Biol. Chem. 277, 883–886 (2002)

    Article  CAS  PubMed  Google Scholar 

  12. K. Alfthan, L. Heiska, M. Gronholm, G.H. Renkema, O. Carpen, Cyclic AMP dependent protein kinase phosphorylates Merlin at serine 518 independently of p21-activated kinase and promotes Merlin-Ezrin heterodimerization. J. Biol. Chem. 279, 1859–1866 (2004)

    Article  Google Scholar 

  13. A.I. McClatchey, Merlin and ERM proteins: unappreciated roles in cancer development? Nat. Rev. Cancer 3, 877–884 (2003)

    Article  PubMed  Google Scholar 

  14. R.J. Shaw, A.I. McClatchey, T. Jacks, Regulation of the neurofibromatosis type 2 tumor suppressor protein, merlin, by adhesion and growth arrest stimuli. J. Biol. Chem. 273, 7757–7764 (1998)

    Article  CAS  PubMed  Google Scholar 

  15. R.J. Shaw, J.G. Paez, M. Curto, A. Yaktine, W.M. Pruitt, I. Saotome, J. O’Bryan, V. Gupta, N. Ratner, C.J. Der, T. Jacks, A.I. McClatchey, The NF2 tumor suppressor, merlin, functions in Rac-dependent signaling. Dev. Cell 1, 63–72 (2001)

    Article  CAS  PubMed  Google Scholar 

  16. T. Okada, M. Lopez-Lago, F.G. Giancotti, Merlin/NF2 mediates contact inhibition of growh by suppressing recruitment of Rac to the plasma membrane. J. Cell Biol. 171, 361–371 (2005)

    Article  CAS  PubMed  Google Scholar 

  17. D. Lallemand, J. Manent, A. Couvelard, A. Watilliaux, M. Siena, F. Chareyre, A. Lampin, M. Niwa-Kawakita, M. Kalamarides, M. Giovannini, Merlin regulates transmembrane receptor accumulation and signaling at the plasma membrane in primary mouse Schwann cells and in human schwannomas. Oncogene 28, 854–865 (2009)

    Article  CAS  PubMed  Google Scholar 

  18. M. Curto, A.I. McClatchey, Nf2/Merlin: A coordinator of receptor signalling and intercellular contact. Br. J. Cancer 98, 256–262 (2008)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. M. Curto, B.K. Cole, D. Lallemand, C.H. Liu, A.I. McClatchey, Contact-dependent inhibition of EGFR signaling by Nf2/Merlin. J. Cell Biol. 177, 893–903 (2007)

    Article  CAS  PubMed  Google Scholar 

  20. A.I. McClatchey, R.G. Fehon, Merlin and the ERM proteins: regulators of receptor distribution and signaling at the cell cortex. Trends Cell Biol. 19, 198–206 (2009)

    Article  PubMed  PubMed Central  Google Scholar 

  21. E.E. Bosco, Y. Nakai, E.F. Hennigan, N. Ratner, Y. Zheng, NF2-deficient cells depend on the Rac1-canonical Wnt signaling pathway to promote the loss of contact inhibition of proliferation. Oncogene 29, 2540–2549 (2010)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. L. Zhou, E. Ercolano, S. Ammoun, M.C. Schmid, M.A. Barczyk, C.O. Hanemann, Merlin-deficient human tumors show loss of contact inhibition and activation of Wnt/β-Catenin signaling linked to the PDGFR/Src and Rac/PAK pathways. Neoplasia 13, 1101–1112 (2011)

    CAS  PubMed  PubMed Central  Google Scholar 

  23. R. Spaventi, L. Pecur, K. Pavelic, Z.P. Pavelic, S. Spaventi, P.J. Stambrook, Human tumour bank in Croatia: a possible model for a small bank as a part of future European tumour bank network. Eur. J. Cancer 30A, 419 (1994)

    Article  CAS  PubMed  Google Scholar 

  24. P. Legoix, M.-F. Legrand, E. Ollagnon, G. Lenoir, G. Thomas, J. Zucman-Rossi, Characterisation of 16 polymorphic markers in the NF2 gene: application to hemizygosity detection. Hum. Mutat. 13, 290–293 (1999)

    Article  CAS  PubMed  Google Scholar 

  25. T. Cacev, S. Radosevic, R. Spaventi, K. Pavelic, S. Kapitanovic, NF1 gene loss of heterozygosity and expression analysis in sporadic colon cancer. Gut 54, 1129–1135 (2005)

    Article  CAS  PubMed  Google Scholar 

  26. D. Hanahan, R.A. Weinberg, Hallmarks of cancer: The next generation. Cell 144, 646–674 (2011)

    Article  CAS  PubMed  Google Scholar 

  27. A.I. McClatchey, I. Saotome, K. Mercer, D. Crowley, J.F. Gusella, R.T. Bronson, T. Jacks, Mice heterozygous for a mutation at the Nf2 tumor suppressor locus develop a range of highly metastatic tumors. Genes Dev. 12, 1121–1133 (1998)

    Article  CAS  PubMed  Google Scholar 

  28. Y.K. Lau, L.B. Murray, S.S. Houshmandi, Y. Xu, D.H. Gutmann, Q. Yu, Merlin is a potent inhibitor of glioma growth. Cancer Res. 68, 5733–5742 (2008)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. G.A. Rouleau, P. Merel, M. Lutchman, M. Sanson, J. Zucman, C. Marineau, K. Hoang-Xuan, S. Demczuk, C. Desmaze, B. Plougastel, Alteration in a new gene encoding a putative membrane-organizing protein causes neurofibromatosis type 2. Nature 363, 515–521 (1993)

    Article  CAS  PubMed  Google Scholar 

  30. A.K. Rustgi, L. Xu, D. Pinney, C. Sterner, R. Beauchamp, S. Schmidt, J.F. Gusella, V. Ramesh, Neurofibromatosis 2 gene in human colorectal cancer. Cancer Genet. Cytogenet. 84, 24–26 (1995)

    Article  CAS  PubMed  Google Scholar 

  31. C.Z. Zhou, Z.H. Peng, F. Zhang, G.Q. Qiu, L. He, Loss of heterozygosity on long arm of chromosome 22 in sporadic colorectal carcinoma. World J. Gastroenterol. 8, 668–673 (2000)

    Google Scholar 

  32. D. Hanahan, R.A. Weinberg, The hallmarks of cancer. Cell 100, 57–70 (2000)

    Article  CAS  PubMed  Google Scholar 

  33. K. Miura, W. Fujibuchi, I. Sasaki, Alternative pre-mRNA splicing in digestive tract malignancy. Cancer Sci. 102, 309–316 (2011)

    Article  CAS  PubMed  Google Scholar 

  34. H. Arakawa, N. Hayashi, H. Nagase, M. Ogawa, Y. Nakamura, Alternative splicing of the NF2 gene and its mutation analysis of breast and colorectal cancers. Hum. Mol. Genet. 3, 565–568 (1994)

    Article  CAS  PubMed  Google Scholar 

  35. T. Sugai, W. Habano, S. Nakamura, N. Uesugi, S. Sasou, C. Itoh, A unique method for mutation analysis of tumor suppressor genes in colorectal carcinomas using a crypt isolation technique. Arch. Pathol. Lab. Med. 124, 382–386 (2000)

    CAS  PubMed  Google Scholar 

  36. N.J. Yoo, S.W. Park, S.H. Lee, Mutational analysis of tumour suppressor gene NF2 in common solid cancers and acute leukaemias. Pathology 44, 29–32 (2012)

    Article  CAS  PubMed  Google Scholar 

Download references

Disclosure/conflict of interest

All experiments were performed in full agreement with ethics, Croatian laws and international conventions. This work was supported by grant number 098-0982464-2508, from the Ministry of Science and Technology, Republic of Croatia. We confirm that we are the sole authors of the contribution which is our original work. It has not been previously published in whole or substantial part and there is no conflict of interest present.

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Authors and Affiliations

  1. Division of Molecular Medicine, Ruđer Bošković Institute, Bijenička cesta 54, 10000, Zagreb, Croatia

    Tamara Čačev & Sanja Kapitanović

  2. Clinical Hospital Dubrava, Avenija Gojka Šuška 6, 10000, Zagreb, Croatia

    Gorana Aralica & Božo Lončar

Authors
  1. Tamara Čačev
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  2. Gorana Aralica
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Correspondence to Tamara Čačev.

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Supplementary Fig. 1

Merlin expression according to TNM stage. 1, weak staining; 2 moderate staining; 3, strong staining (DOCX 58 kb)

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Čačev, T., Aralica, G., Lončar, B. et al. Loss of NF2/Merlin expression in advanced sporadic colorectal cancer. Cell Oncol. 37, 69–77 (2014). https://doi.org/10.1007/s13402-013-0164-2

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