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The miRNA-200 family and miRNA-9 exhibit differential expression in primary versus corresponding metastatic tissue in breast cancer

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Abstract

Metastases are the major cause of cancer-related deaths, but the mechanisms of the metastatic process remain poorly understood. In recent years, the involvement of microRNAs (miRNAs) in cancer has become apparent, and the objective of this study was to identify miRNAs associated with breast cancer progression. Global miRNA expression profiling was performed on 47 tumor samples from 14 patients with paired samples from primary breast tumors and corresponding lymph node and distant metastases using LNA-enhanced miRNA microarrays. The identified miRNA expression alterations were validated by real-time PCR, and tissue distribution of the miRNAs was visualized by in situ hybridization. The patients, in which the miRNA profile of the primary tumor and corresponding distant metastasis clustered in the unsupervised cluster analysis, showed significantly shorter intervals between the diagnosis of the primary tumor and distant metastasis (median 1.6 years) compared to those that did not cluster (median 11.3 years) (p < 0.003). Fifteen miRNAs were identified that were significantly differentially expressed between primary tumors and corresponding distant metastases, including miR-9, miR-219-5p and four of the five members of the miR-200 family involved in epithelial-mesenchymal transition. Tumor expression of miR-9 and miR-200b were confirmed using in situ hybridization, which also verified higher expression of these miRNAs in the distant metastases versus corresponding primary tumors. Our results demonstrate alterations in miRNA expression at different stages of disease progression in breast cancer, and suggest a direct involvement of the miR-200 family and miR-9 in the metastatic process.

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References

  1. (IARC) IAfRoC (2010) Globocan 2008. http://globocaniarcfr/factsheets/populations/factsheetasp?uno=900#KEY

  2. Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116(2):281–297

    Article  PubMed  CAS  Google Scholar 

  3. Tsuchiya S, Okuno Y, Tsujimoto G (2006) MicroRNA: biogenetic and functional mechanisms and involvements in cell differentiation and cancer. J Pharmacol Sci 101(4):267–270

    Article  PubMed  CAS  Google Scholar 

  4. Lewis BP, Burge CB, Bartel DP (2005) Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 120(1):15–20

    Article  PubMed  CAS  Google Scholar 

  5. Iorio MV, Ferracin M, Liu CG, Veronese A, Spizzo R, Sabbioni S et al (2005) MicroRNA gene expression deregulation in human breast cancer. Cancer Res 65(16):7065–7070

    Article  PubMed  CAS  Google Scholar 

  6. Lu J, Getz G, Miska EA, Alvarez-Saavedra E, Lamb J, Peck D et al (2005) MicroRNA expression profiles classify human cancers. Nature 435(7043):834–838

    Article  PubMed  CAS  Google Scholar 

  7. Rosenfeld N, Aharonov R, Meiri E, Rosenwald S, Spector Y, Zepeniuk M et al (2008) MicroRNAs accurately identify cancer tissue origin. Nat Biotechnol 26(4):462–469

    Article  PubMed  CAS  Google Scholar 

  8. Rosenwald S, Gilad S, Benjamin S, Lebanony D, Dromi N, Faerman A et al (2010) Validation of a microRNA-based qRT-PCR test for accurate identification of tumor tissue origin. Mod Pathol 23(6):814–823

    Article  PubMed  CAS  Google Scholar 

  9. Valastyan S, Reinhardt F, Benaich N, Calogrias D, Szasz AM, Wang ZC et al (2009) A pleiotropically acting microRNA, miR-31, inhibits breast cancer metastasis. Cell 137(6):1032–1046

    Article  PubMed  CAS  Google Scholar 

  10. Tavazoie SF, Alarcon C, Oskarsson T, Padua D, Wang Q, Bos PD et al (2008) Endogenous human microRNAs that suppress breast cancer metastasis. Nature 451(7175):147–152

    Article  PubMed  CAS  Google Scholar 

  11. Zhu S, Wu H, Wu F, Nie D, Sheng S, Mo YY (2008) MicroRNA-21 targets tumor suppressor genes in invasion and metastasis. Cell Res 18(3):350–359

    Article  PubMed  CAS  Google Scholar 

  12. Huang Q, Gumireddy K, Schrier M, le Sage C, Nagel R, Nair S et al (2008) The microRNAs miR-373 and miR-520c promote tumour invasion and metastasis. Nat Cell Biol 10(2):202–210

    Article  PubMed  CAS  Google Scholar 

  13. Griffiths-Jones S, Saini HK, van Dongen S, Enright AJ (2008) miRBase: tools for microRNA genomics. Nucleic Acids Res 36(Database issue):D154–D158

    PubMed  CAS  Google Scholar 

  14. Kozomara A, Griffiths-Jones S (2011) miRBase: integrating microRNA annotation and deep-sequencing data. Nucleic Acids Res 39(Database issue):D152–D157

    Article  PubMed  Google Scholar 

  15. Park SM, Gaur AB, Lengyel E, Peter ME (2008) The miR-200 family determines the epithelial phenotype of cancer cells by targeting the E-cadherin repressors ZEB1 and ZEB2. Genes Dev 22(7):894–907

    Article  PubMed  CAS  Google Scholar 

  16. Gregory PA, Bert AG, Paterson EL, Barry SC, Tsykin A, Farshid G et al (2008) The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1. Nat Cell Biol 10(5):593–601

    Article  PubMed  CAS  Google Scholar 

  17. Birchmeier C, Birchmeier W, Brand-Saberi B (1996) Epithelial-mesenchymal transitions in cancer progression. Acta Anat 156(3):217–226

    Article  PubMed  CAS  Google Scholar 

  18. Chaffer CL, Weinberg RA (2011) A perspective on cancer cell metastasis. Science 331(6024):1559–1564

    Article  PubMed  CAS  Google Scholar 

  19. Korpal M, Lee ES, Hu G, Kang Y (2008) The miR-200 family inhibits epithelial-mesenchymal transition and cancer cell migration by direct targeting of E-cadherin transcriptional repressors ZEB1 and ZEB2. J Biol Chem 283(22):14910–14914

    Article  PubMed  CAS  Google Scholar 

  20. Hugo H, Ackland ML, Blick T, Lawrence MG, Clements JA, Williams ED et al (2007) Epithelial–mesenchymal and mesenchymal–epithelial transitions in carcinoma progression. J Cell Physiol 213(2):374–383

    Article  PubMed  CAS  Google Scholar 

  21. Ma L, Young J, Prabhala H, Pan E, Mestdagh P, Muth D et al (2010) miR-9, a MYC/MYCN-activated microRNA, regulates E-cadherin and cancer metastasis. Nat Cell Biol 12(3):247–256

    PubMed  CAS  Google Scholar 

  22. Zhu L, Chen H, Zhou D, Li D, Bai R, Zheng S, et al. (2011) MicroRNA-9 up-regulation is involved in colorectal cancer metastasis via promoting cell motility. Med Oncol

  23. Laios A, O’Toole S, Flavin R, Martin C, Kelly L, Ring M et al (2008) Potential role of miR-9 and miR-223 in recurrent ovarian cancer. Mol Cancer 7:35

    Article  PubMed  Google Scholar 

  24. Hu X, Schwarz JK, Lewis JS Jr, Huettner PC, Rader JS, Deasy JO et al (2010) A microRNA expression signature for cervical cancer prognosis. Cancer Res 70(4):1441–1448

    Article  PubMed  CAS  Google Scholar 

  25. Tan HX, Wang Q, Chen LZ, Huang XH, Chen JS, Fu XH et al (2010) MicroRNA-9 reduces cell invasion and E-cadherin secretion in SK-Hep-1 cell. Med Oncol 27(3):654–660

    Article  PubMed  CAS  Google Scholar 

  26. Guo LM, Pu Y, Han Z, Liu T, Li YX, Liu M et al (2009) MicroRNA-9 inhibits ovarian cancer cell growth through regulation of NF-kappaB1. FEBS J 276(19):5537–5546

    Article  PubMed  CAS  Google Scholar 

  27. Wan HY, Guo LM, Liu T, Liu M, Li X, Tang H (2010) Regulation of the transcription factor NF-kappaB1 by microRNA-9 in human gastric adenocarcinoma. Mol Cancer 9:16

    Article  PubMed  Google Scholar 

  28. Benjamini Y, Hochberg Y (1995) Controlling the false discovery rate: a practical and powerful approach to multiple testing. J Roy Statist 57(1):289–300

    Google Scholar 

  29. Schopman NC, Heynen S, Haasnoot J, Berkhout B (2010) A miRNA-tRNA mix-up: tRNA origin of proposed miRNA. RNA Biol 7(5):573–576

    Article  PubMed  CAS  Google Scholar 

  30. Korpal M, Kang Y (2008) The emerging role of miR-200 family of microRNAs in epithelial-mesenchymal transition and cancer metastasis. RNA Biol 5(3):115–119

    Article  PubMed  CAS  Google Scholar 

  31. Fidler IJ, Kripke ML (1977) Metastasis results from preexisting variant cells within a malignant tumor. Science 197(4306):893–895

    Article  PubMed  CAS  Google Scholar 

  32. van ‘t Veer LJ, Dai H, van de Vijver MJ, He YD, Hart AA, Mao M et al (2002) Gene expression profiling predicts clinical outcome of breast cancer. Nature 415(6871):530–536

    Article  Google Scholar 

  33. van de Vijver MJ, He YD, van’t Veer LJ, Dai H, Hart AA, Voskuil DW et al (2002) A gene-expression signature as a predictor of survival in breast cancer. N Engl J Med 347(25):1999–2009

    Article  PubMed  Google Scholar 

  34. Kowalski PJ, Rubin MA, Kleer CG (2003) E-cadherin expression in primary carcinomas of the breast and its distant metastases. Breast Cancer Res 5(6):R217–R222

    Article  PubMed  CAS  Google Scholar 

  35. Nass SJ, Herman JG, Gabrielson E, Iversen PW, Parl FF, Davidson NE et al (2000) Aberrant methylation of the estrogen receptor and E-cadherin 5′ CpG islands increases with malignant progression in human breast cancer. Cancer Res 60(16):4346–4348

    PubMed  CAS  Google Scholar 

  36. Younis LK, El Sakka H, Haque I (2007) The prognostic value of E-cadherin expression in breast cancer. Int J Health Sci 1(1):43–51

    Google Scholar 

  37. Hwang HW, Wentzel EA, Mendell JT (2007) A hexanucleotide element directs microRNA nuclear import. Science 315(5808):97–100

    Article  PubMed  CAS  Google Scholar 

  38. Jeffries CD, Fried HM, Perkins DO (2011) Nuclear and cytoplasmic localization of neural stem cell microRNAs. RNA 17(4):675–686

    Article  PubMed  CAS  Google Scholar 

  39. Nass D, Rosenwald S, Meiri E, Gilad S, Tabibian-Keissar H, Schlosberg A et al (2009) MiR-92b and miR-9/9* are specifically expressed in brain primary tumors and can be used to differentiate primary from metastatic brain tumors. Brain Pathol 19(3):375–383

    Article  PubMed  CAS  Google Scholar 

  40. Sempere LF, Freemantle S, Pitha-Rowe I, Moss E, Dmitrovsky E, Ambros V (2004) Expression profiling of mammalian microRNAs uncovers a subset of brain-expressed microRNAs with possible roles in murine and human neuronal differentiation. Genome Biol 5(3):R13

    Article  PubMed  Google Scholar 

  41. Krichevsky AM, King KS, Donahue CP, Khrapko K, Kosik KS (2003) A microRNA array reveals extensive regulation of microRNAs during brain development. RNA 9(10):1274–1281

    Article  PubMed  CAS  Google Scholar 

  42. Lukiw WJ (2007) Micro-RNA speciation in fetal, adult and Alzheimer’s disease hippocampus. Neuroreport 18(3):297–300

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We thank Lisbeth Mortensen and Ole Nielsen for excellent technical assistance with the immunohistochemical analysis and M. K. Occhipinti-Bender for editorial assistance. This study was supported in part by the Danish Cancer Society, the Danish Cancer Research Foundation, A Race Against Breast Cancer, Sino-Danish Breast Cancer Research Centre, NanoCAN Lundbeck Center of Excellence, and Danish Center for Translational Breast Cancer research (DCTB).

Ethical Standards

This manuscript complies with the currents laws in Denmark.

Conflict of interest

BSN is employee at Bioneer A/S. RS and TL are former employees of Exiqon A/S. All other authors declare that they have no conflict of interest.

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Author notes

  1. Boye Schnack Nielsen

    Present address: Bioneer A/S, Horsholm, Denmark

Authors and Affiliations

  1. Department of Cancer and Inflammation Research, Institute of Molecular Medicine, University of Southern Denmark, J. B. Winsloewsvej 25, 5000, Odense C, Denmark

    Karina H. Gravgaard, Maria B. Lyng & Henrik J. Ditzel

  2. Department of Pathology, Hospital South, Slagelse, Denmark

    Anne-Vibeke Laenkholm

  3. Department of Biomarker Discovery, Exiqon A/S, Vedbaek, Denmark

    Rolf Søkilde, Boye Schnack Nielsen & Thomas Litman

  4. Department of Oncology, Odense University Hospital, Odense, Denmark

    Henrik J. Ditzel

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  1. Karina H. Gravgaard
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Correspondence to Henrik J. Ditzel.

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Gravgaard, K.H., Lyng, M.B., Laenkholm, AV. et al. The miRNA-200 family and miRNA-9 exhibit differential expression in primary versus corresponding metastatic tissue in breast cancer. Breast Cancer Res Treat 134, 207–217 (2012). https://doi.org/10.1007/s10549-012-1969-9

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  • DOI: https://doi.org/10.1007/s10549-012-1969-9

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