Advertisement

Breast Cancer

, Volume 21, Issue 6, pp 715–723 | Cite as

Collapsin response mediator protein 2 is involved in regulating breast cancer progression

  • Kazuhiro ShimadaEmail author
  • Takashi Ishikawa
  • Fumio Nakamura
  • Daisuke Shimizu
  • Takashi Chishima
  • Yasushi Ichikawa
  • Takeshi Sasaki
  • Itaru Endo
  • Yoji Nagashima
  • Yoshio Goshima
Original Article

Abstract

Background

Altered expression of collapsin response mediator proteins (CRMPs) has been reported in several malignant tumors, including downregulation of CRMP1 in lung cancer and upregulation of CRMP2 in colorectal cancer. This study aimed to investigate the relationship between CRMP expression and clinicopathological characteristics in patients with breast cancer.

Methods

Twenty-two breast cancer and four normal breast tissues were used to assess CRMP mRNA expression. The average expression level of each CRMP (CRMP15) mRNA was analyzed in a subset of breast cancer specimens and compared with that in normal breast tissue by real-time quantitative reverse-transcription polymerase chain reaction. Furthermore, 173 breast cancer specimens and matching normal breast controls were used for immunohistochemistry based on the tissue microarray technique. Levels of CRMP2 and phosphorylated CRMP2 protein were assessed, and possible correlations between the clinicopathological characteristics were evaluated.

Results:

The expression of CRMP2 mRNA was significantly decreased in breast cancer tissues, while that of the other CRMPs was similar between normal and breast cancer tissues. Immunohistochemistry revealed that CRMP2 protein expression was also decreased in breast cancer tissues (P < 0.001). Phosphorylated CRMP2 was observed in the nuclei of breast cancer cells but not in normal mammary cells (P < 0.001). Furthermore, nuclear phosphorylated CRMP2 expression was increased in proportion to the histological grade and triple-negative subtype.

Conclusions

Reduced CRMP2 expression and elevated expression of nuclear phosphorylated CRMP2 may be associated with breast cancer progression.

Keywords

Breast cancer Collapsin response mediator protein 2 Nuclear localization Microtubule Triple-negative breast cancer 

Notes

Acknowledgments

We thank Msato Kodera, Harumi Sakurada, Yuka Honjoh, Makiko Fujimura and Yumi Inada for their technical and secretarial assistance. We also thank the staff of the Human Cancer Tissue Center of Kanagawa Cancer Research and Information Association for preparation of tissue samples and clinicopathological information. This study was supported by Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science under the Ministry of Education, Culture, Sports, Science and Technology (Fundamental Research: C, 23591899) (to Takashi Ishikawa, no. 80275049).

Conflict of interest

The authors declare that they have no conflict of interests.

References

  1. 1.
    DeSantis C, Siegel R, Bandi P, Jemal A. Breast cancer statistics. CA Cancer J Clin. 2011;61:409–18.CrossRefPubMedGoogle Scholar
  2. 2.
    Paik S, Shak S, Tang G, Kim C, Baker J, Cronin M, et al. A multigene assay to predict recurrence of tamoxifen-treated, node-negative breast cancer. N Engl J Med. 2004;351:2817–26.CrossRefPubMedGoogle Scholar
  3. 3.
    Paik S, Tang G, Shak S, Kim C, Baker J, Kim W, et al. Gene expression and benefit of chemotherapy in women with node-negative, estrogen receptor-positive breast cancer. J Clin Oncol. 2006;24:3726–34.CrossRefPubMedGoogle Scholar
  4. 4.
    Goshima Y, Nakamura F, Strittmatter P, Strittmatter SM. Collapsin-induced growth cone collapse mediated by an intracellular protein related to UNC-33. Nature. 1995;376:509–14.CrossRefPubMedGoogle Scholar
  5. 5.
    Minturn JE, Fryer HJ, Geschwind DH, Hockfield S. TOAD-64, a gene expressed early in neuronal differentiation in the rat, is related to unc-33, a C. elegans gene involved in axon outgrowth. J Neurosci. 1995;15:6757–66.PubMedGoogle Scholar
  6. 6.
    Arimura N, Inagaki N, Chihara K, Menager C, Nakamura N, Amano M, et al. Phosphorylation of collapsin response mediator protein-2 by Rho-kinase. Evidence for two separate signaling pathways for growth cone collapse. J Biol Chem. 2000;275:23973–80.CrossRefPubMedGoogle Scholar
  7. 7.
    Schmidt EF, Strittmatter SM. The CRMP family of proteins and their role in Sema3A signaling. Adv Exp Med Biol. 2007;600:1–11.CrossRefPubMedCentralPubMedGoogle Scholar
  8. 8.
    Wang LH, Strittmatter SM. Brain CRMP forms heterotetramers similar to liver dihydropyrimidinase. J Neurochem. 1997;69:2261–9.CrossRefPubMedGoogle Scholar
  9. 9.
    Fukada M, Watakabe I, Yuasa-Kawada J, Kawachi H, Kuroiwa A, Matsuda Y, et al. Molecular characterization of CRMP5, a novel member of the collapsin response mediator protein family. J Biol Chem. 2000;275:37957–65.CrossRefPubMedGoogle Scholar
  10. 10.
    Gaetano C, Matsuo T, Thiele CJ. Identification and characterization of a retinoic acid-regulated human homologue of the unc-33-like phosphoprotein gene (hUlip) from neuroblastoma cells. J Biol Chem. 1997;272:12195–201.CrossRefPubMedGoogle Scholar
  11. 11.
    Nishimura T, Fukata Y, Kato K, Yamaguchi T, Matsuura Y, Kamiguchi H, et al. CRMP-2 regulates polarized Numb-mediated endocytosis for axon growth. Nat Cell Biol. 2003;5:819–26.CrossRefPubMedGoogle Scholar
  12. 12.
    Kimura T, Watanabe H, Iwamatsu A, Kaibuchi K. Tubulin and CRMP-2 complex is transported via Kinesin-1. J Neurochem. 2005;93:1371–82.CrossRefPubMedGoogle Scholar
  13. 13.
    Fukata Y, Itoh TJ, Kimura T, Menager C, Nishimura T, Shiromizu T, et al. CRMP-2 binds to tubulin heterodimers to promote microtubule assembly. Nat Cell Biol. 2002;4:583–91.PubMedGoogle Scholar
  14. 14.
    Brown M, Jacobs T, Eickholt B, Ferrari G, Teo M, Monfries C, et al. Alpha2-chimaerin, cyclin-dependent Kinase 5/p35, and its target collapsin response mediator protein-2 are essential components in semaphorin 3A-induced growth-cone collapse. J Neurosci. 2004;24:8994–9004.CrossRefPubMedGoogle Scholar
  15. 15.
    Sasaki Y, Cheng C, Uchida Y, Nakajima O, Ohshima T, Yagi T, et al. Fyn and Cdk5 mediate semaphorin-3A signaling, which is involved in regulation of dendrite orientation in cerebral cortex. Neuron. 2002;35:907–20.CrossRefPubMedGoogle Scholar
  16. 16.
    Uchida Y, Ohshima T, Sasaki Y, Suzuki H, Yanai S, Yamashita N, et al. Semaphorin3A signalling is mediated via sequential Cdk5 and GSK3beta phosphorylation of CRMP2: implication of common phosphorylating mechanism underlying axon guidance and Alzheimer’s disease. Genes Cells. 2005;10:165–79.CrossRefPubMedGoogle Scholar
  17. 17.
    Cole AR, Knebel A, Morrice NA, Robertson LA, Irving AJ, Connolly CN, et al. GSK-3 phosphorylation of the Alzheimer epitope within collapsin response mediator proteins regulates axon elongation in primary neurons. J Biol Chem. 2004;279:50176–80.CrossRefPubMedCentralPubMedGoogle Scholar
  18. 18.
    Inagaki N, Chihara K, Arimura N, Menager C, Kawano Y, Matsuo N, et al. CRMP-2 induces axons in cultured hippocampal neurons. Nat Neurosci. 2001;4:781–2.CrossRefPubMedGoogle Scholar
  19. 19.
    Yoshimura T, Kawano Y, Arimura N, Kawabata S, Kikuchi A, Kaibuchi K. GSK-3beta regulates phosphorylation of CRMP-2 and neuronal polarity. Cell. 2005;120:137–49.CrossRefPubMedGoogle Scholar
  20. 20.
    Lin PC, Chan PM, Hall C, Manser E. Collapsin response mediator proteins (CRMPs) are a new class of microtubule-associated protein (MAP) that selectively interacts with assembled microtubules via a taxol-sensitive binding interaction. J Biol Chem. 2011;286:41466–78.CrossRefPubMedCentralPubMedGoogle Scholar
  21. 21.
    Gu Y, Hamajima N, Ihara Y. Neurofibrillary tangle-associated collapsin response mediator protein-2 (CRMP-2) is highly phosphorylated on Thr-509, Ser-518, and Ser-522. Biochemistry. 2000;39:4267–75.CrossRefPubMedGoogle Scholar
  22. 22.
    Yoshida H, Watanabe A, Ihara Y. Collapsin response mediator protein-2 is associated with neurofibrillary tangles in Alzheimer’s disease. J Biol Chem. 1998;273:9761–8.CrossRefPubMedGoogle Scholar
  23. 23.
    Cole AR, Noble W, van Aalten L, Plattner F, Meimaridou R, Hogan D, et al. Collapsin response mediator protein-2 hyperphosphorylation is an early event in Alzheimer’s disease progression. J Neurochem. 2007;103:1132–44.CrossRefPubMedGoogle Scholar
  24. 24.
    Soutar MP, Thornhill P, Cole AR, Sutherland C. Increased CRMP2 phosphorylation is observed in Alzheimer’s disease; does this tell us anything about disease development? Curr Alzheimer Res. 2009;6:269–78.CrossRefPubMedGoogle Scholar
  25. 25.
    Wu CC, Chen HC, Chen SJ, Liu HP, Hsieh YY, Yu CJ, et al. Identification of collapsin response mediator protein-2 as a potential marker of colorectal carcinoma by comparative analysis of cancer cell secretomes. Proteomics. 2008;8:316–32.CrossRefPubMedGoogle Scholar
  26. 26.
    Shih JY, Yang SC, Hong TM, Yuan A, Chen JJ, Yu CJ, et al. Collapsin response mediator protein-1 and the invasion and metastasis of cancer cells. J Natl Cancer Inst. 2001;93:1392–400.CrossRefPubMedGoogle Scholar
  27. 27.
    Gao X, Pang J, Li LY, Liu WP, Di JM, Sun QP, et al. Expression profiling identifies new function of collapsin response mediator protein 4 as a metastasis-suppressor in prostate cancer. Oncogene. 2010;29:4555–66.CrossRefPubMedGoogle Scholar
  28. 28.
    Lakhani SR, Ellis IO, Schnitt SJ, Tan PH, van de Vijver MJ. WHO classification of tumours of the breast. 4th edn. World Health Organization; 2012.Google Scholar
  29. 29.
    Elston CW, Ellis IO. Pathological prognostic factors in breast cancer. I. The value of histological grade in breast cancer: experience from a large study with long-term follow-up. Histopathology. 1991;19:403–10.CrossRefPubMedGoogle Scholar
  30. 30.
    Kojima Y, Akimoto K, Nagashima Y, Ishiguro H, Shirai S, Chishima T, et al. The overexpression and altered localization of the atypical protein kinase C lambda/iota in breast cancer correlates with the pathologic type of these tumors. Hum Pathol. 2008;39:824–31.CrossRefPubMedGoogle Scholar
  31. 31.
    Takagawa R, Akimoto K, Ichikawa Y, Akiyama H, Kojima Y, Ishiguro H, et al. High expression of atypical protein kinase C lambda/iota in gastric cancer as a prognostic factor for recurrence. Ann Surg Oncol. 2010;17:81–8.CrossRefPubMedGoogle Scholar
  32. 32.
    Goodyear S, Sharma MC. Roscovitine regulates invasive breast cancer cell (MDA-MB231) proliferation and survival through cell cycle regulatory protein cdk5. Exp Mol Pathol. 2007;82:25–32.CrossRefPubMedGoogle Scholar
  33. 33.
    Upadhyay AK, Ajay AK, Singh S, Bhat MK. Cell cycle regulatory protein 5 (Cdk5) is a novel downstream target of ERK in carboplatin induced death of breast cancer cells. Curr Cancer Drug Targets. 2008;8:741–52.CrossRefPubMedGoogle Scholar
  34. 34.
    Prasad CP, Rath G, Mathur S, Bhatnagar D, Parshad R, Ralhan R. Expression analysis of E-cadherin, Slug and GSK3beta in invasive ductal carcinoma of breast. BMC Cancer. 2009;9:325.CrossRefPubMedCentralPubMedGoogle Scholar
  35. 35.
    Nowak AK, Wilcken NR, Stockler MR, Hamilton A, Ghersi D. Systematic review of taxane-containing versus non-taxane-containing regimens for adjuvant and neoadjuvant treatment of early breast cancer. Lancet Oncol. 2004;5:372–80.CrossRefPubMedGoogle Scholar
  36. 36.
    Trudeau M, Charbonneau F, Gelmon K, Laing K, Latreille J, Mackey J, et al. Selection of adjuvant chemotherapy for treatment of node-positive breast cancer. Lancet Oncol. 2005;6:886–98.CrossRefPubMedGoogle Scholar
  37. 37.
    Amos LA, Lowe J. How taxol stabilises microtubule structure. Chem Biol. 1999;6:R65–9.CrossRefPubMedGoogle Scholar
  38. 38.
    Burkhart CA, Kavallaris M, Band Horwitz M. The role of beta-tubulin isotypes in resistance to antimitotic drugs. Biochim Biophys Acta. 2001;1471:O1–9.PubMedGoogle Scholar
  39. 39.
    Cleator S, Heller W, Coombes RC. Triple-negative breast cancer: therapeutic options. Lancet Oncol. 2007;8:235–44.CrossRefPubMedGoogle Scholar
  40. 40.
    Miyoshi Y, Kurosumi M, Kurebayashi J, Matsuura N, Takahashi M, Tokunaga E, et al. Low nuclear grade but not cell proliferation predictive of pathological complete response to docetaxel in human breast cancers. J Cancer Res Clin Oncol. 2008;134:561–7.CrossRefPubMedGoogle Scholar
  41. 41.
    Paradiso A, Mangia A, Chiriatti A, Tommasi S, Zito A, Latorre A, et al. Biomarkers predictive for clinical efficacy of taxol-based chemotherapy in advanced breast cancer. Ann Oncol. 2005;16 Suppl 4:iv14–19.Google Scholar
  42. 42.
    Tommasi S, Mangia A, Lacalamita R, Bellizzi A, Fedele V, Chiriatti A, et al. Cytoskeleton and paclitaxel sensitivity in breast cancer: the role of beta-tubulins. Int J Cancer. 2007;120:2078–85.CrossRefPubMedGoogle Scholar
  43. 43.
    Wagner P, Wang B, Clark E, Lee H, Rouzier R, Pusztai L. Microtubule associated protein (MAP)-tau: a novel mediator of paclitaxel sensitivity in vitro and in vivo. Cell Cycle. 2005;4:1149–52.CrossRefPubMedGoogle Scholar
  44. 44.
    Rouzier R, Rajan R, Wagner P, Hess KR, Gold DL, Stec J, et al. Microtubule-associated protein tau: a marker of paclitaxel sensitivity in breast cancer. Proc Natl Acad Sci USA. 2005;102:8315–20.CrossRefPubMedCentralPubMedGoogle Scholar
  45. 45.
    Hasegawa S, Miyoshi Y, Egawa C, Ishitobi M, Taguchi T, Tamaki Y, et al. Prediction of response to docetaxel by quantitative analysis of class I and III beta-tubulin isotype mRNA expression in human breast cancers. Clin Cancer Res. 2003;9:2992–7.PubMedGoogle Scholar
  46. 46.
    Smoter M, Bodnar L, Duchnowska R, Stec R, Grala B, Szczylik C. The role of Tau protein in resistance to paclitaxel. Cancer Chemother Pharmacol. 2011;68:553–7.CrossRefPubMedCentralPubMedGoogle Scholar

Copyright information

© The Japanese Breast Cancer Society 2013

Authors and Affiliations

  • Kazuhiro Shimada
    • 1
    • 2
    Email author
  • Takashi Ishikawa
    • 2
  • Fumio Nakamura
    • 3
  • Daisuke Shimizu
    • 2
  • Takashi Chishima
    • 1
    • 4
  • Yasushi Ichikawa
    • 1
    • 4
  • Takeshi Sasaki
    • 5
  • Itaru Endo
    • 1
    • 4
  • Yoji Nagashima
    • 6
  • Yoshio Goshima
    • 3
  1. 1.Department of Gastroenterological SurgeryYokohama City University Graduate School of MedicineYokohamaJapan
  2. 2.Department of Breast and Thyroid SurgeryYokohama City University Medical CenterYokohamaJapan
  3. 3.Department of Molecular Pharmacology and NeurobiologyYokohama City University Graduate School of MedicineYokohamaJapan
  4. 4.Department of Clinical OncologyYokohama City University Graduate School of MedicineYokohamaJapan
  5. 5.Division of Surgical PathologyYokohama City University Medical CenterYokohamaJapan
  6. 6.Department of Molecular PathologyYokohama City University Graduate School of MedicineYokohamaJapan

Personalised recommendations