Differential expression of miR-139, miR-486 and miR-21 in breast cancer patients sub-classified according to lymph node status
- 526 Downloads
Therapeutic decisions in breast cancer are increasingly guided by prognostic and predictive biomarkers. Non-protein-coding microRNAs (miRNAs) have recently been found to be deregulated in breast cancers and, in addition, to be correlated with several clinico-pathological features. One of the most consistently up-regulated miRNAs is miR-21. Here, we specifically searched for differentially expressed miRNAs in high-risk breast cancer patients as compared to low-risk breast cancer patients. In the same patients, we also compared miR-21 expression with the expression of its presumed target PTEN.
Both microarray and RT-qPCR techniques were used to assess miRNA expression levels in lymph node-positive and -negative human invasive ductal carcinoma tissues. Simultaneously, PTEN protein expression levels were assessed using immunohistochemistry.
miR-486-5p and miR-139-5p were found to be down-regulated in patients with lymph node metastases, whereas miR-21 was found to be up-regulated in patients with a positive lymph node status. miR-21 expression levels were found to significantly correlate with tumour size (r = 0.403, p = 0.009; Spearman’s rank), whereas no relation was found between miR-21 and PTEN expression levels (Kruskal-Wallis test).
Down-regulation of miR-486-5p and miR-139-5p, in conjunction with up-regulation of miR-21, may represent a useful signature for the identification of high-risk breast cancer patients.
KeywordsBreast cancer miRNA miR-21 miR-486 miR-139 Biomarker
We thank Stine Jørgensen and Jette Nyvang Jørgensen for their help with patient records and tissue sample collections, as well as Mel Charles John Heeran for skilful PTEN staining of the tissue sections. Birgit Hertz, Marianne Fregil and Gitte Friis are acknowledged for their excellent technical assistance in the laboratory. Moreover, we thank Søren Møller and Exiqon for performing the microarray experiments. We are grateful to Catherine Aubel for her careful and professional proofreading of the manuscript. Furthermore, we are indebted to Susanne Husted and Boye Schnack Nielsen for qPCR discussion and antibody selection, respectively. We thank the anonymous Cellular Oncology referees for constructive comments and comprehensive proofreading. The Danish CancerBiobank (DCB) and the MAMBIO breast carcinoma biobank are acknowledged for biological material and for information regarding handling and storage. The work was funded by the Danish Graduate School in Clinical Oncology and the Research Foundation of Copenhagen County.
Conflict of interest
During the experiments performed for this manuscript Thomas Litman and Rolf Søkilde were employed full-time for an entity (Exiqon A/S) having a commercial interest in the microarray platform used.
- 7.I.A. Asangani, S.A. Rasheed, D.A. Nikolova, J.H. Leupold, N.H. Colburn, S. Post, H. Allgayer, MicroRNA-21 (miR-21) post-transcriptionally downregulates tumour suppressor Pdcd4 and stimulates invasion, intravasation and metastasis in colorectal cancer. Oncogene 27, 2128–2136 (2008)PubMedCrossRefGoogle Scholar
- 8.H.K. Oh, A.L. Tan, K. Das, C.H. Ooi, N.T. Deng, I.B. Tan, E. Beillard, J. Lee, K. Ramnarayanan, S.Y. Rha, N. Palanisamy, P.M. Voorhoeve, P. Tan, Genomic loss of miR-486 regulates tumour progression and the OLFM4 antiapoptotic factor in gastric cancer. Clin Cancer Res 17, 2657–2667 (2011)PubMedCrossRefGoogle Scholar
- 11.M.V. Iorio, M. Ferracin, C.G. Liu, A. Veronese, R. Spizzo, S. Sabbioni, E. Magri, M. Pedriali, M. Fabbri, M. Campiglio, S. Menard, J.P. Palazzo, A. Rosenberg, P. Musiani, S. Volinia, I. Nenci, G.A. Calin, P. Querzoli, M. Negrini, C.M. Croce, MicroRNA gene expression deregulation in human breast cancer. Cancer Res 65, 7065–7070 (2005)PubMedCrossRefGoogle Scholar
- 13.M.D. Mattie, C.C. Benz, J. Bowers, K. Sensinger, L. Wong, G.K. Scott, V. Fedele, D. Ginzinger, R. Getts, C. Haqq, Optimized high-throughput microRNA expression profiling provides novel biomarker assessment of clinical prostate and breast cancer biopsies. Mol Cancer 5, 24 (2006)PubMedCentralPubMedCrossRefGoogle Scholar
- 14.A.J. Lowery, N. Miller, A. Devaney, R.E. McNeill, P.A. Davoren, C. Lemetre, V. Benes, S. Schmidt, J. Blake, G. Ball, M.J. Kerin, MicroRNA signatures predict oestrogen receptor, progesterone receptor and HER2/neu receptor status in breast cancer. Breast Cancer Res 11, R27 (2009)PubMedCentralPubMedCrossRefGoogle Scholar
- 17.L.X. Yan, Q.N. Wu, Y. Zhang, Y.Y. Li, D.Z. Liao, J.H. Hou, J. Fu, M.S. Zeng, J.P. Yun, Q.L. Wu, Y.X. Zeng, J.Y. Shao, Knockdown of miR-21 in human breast cancer cell lines inhibits proliferation, in vitro migration and in vivo tumour growth. Breast Cancer Res 13, R2 (2011)PubMedCentralPubMedCrossRefGoogle Scholar
- 27.A. Perren, L.P. Weng, A.H. Boag, U. Ziebold, K. Thakore, P.L. Dahia, P. Komminoth, J.A. Lees, L.M. Mulligan, G.L. Mutter, C. Eng, Immunohistochemical evidence of loss of PTEN expression in primary ductal adenocarcinomas of the breast. Am J Pathol 155, 1253–1260 (1999)PubMedCentralPubMedCrossRefGoogle Scholar
- 38.A. Smeets, A. Daemen, B. Vanden, I, O. Gevaert, B. Claes, H. Wildiers, R. Drijkoningen, H. P. Van, D. Lambrechts, M. B. De, P. Neven, C. Sotiriou, T. Vandorpe, R. Paridaens and M. R. Christiaens, Prediction of lymph node involvement in breast cancer from primary tumour tissue using gene expression profiling and miRNAs. Breast Cancer Res Treat 129, 767–776 (2011)PubMedCrossRefGoogle Scholar
- 45.S. Corbetta, V. Vaira, V. Guarnieri, A. Scillitani, C. Eller-Vainicher, S. Ferrero, L. Vicentini, I. Chiodini, M. Bisceglia, P. Beck-Peccoz, S. Bosari, A. Spada, Differential expression of microRNAs in human parathyroid carcinomas compared with normal parathyroid tissue. Endocr Relat Cancer 17, 135–146 (2010)PubMedCrossRefGoogle Scholar
- 51.W. Bao, H.J. Fu, Q.S. Xie, L. Wang, R. Zhang, Z.Y. Guo, J. Zhao, Y.L. Meng, X.L. Ren, T. Wang, Q. Li, B.Q. Jin, L.B. Yao, R.A. Wang, D.M. Fan, S.Y. Chen, L.T. Jia, A.G. Yang, HER2 interacts with CD44 to up-regulate CXCR4 via epigenetic silencing of microRNA-139 in gastric cancer cells. Gastroenterology 141, 2076–2087 (2011)PubMedCrossRefGoogle Scholar
- 55.L.F. Sempere, M. Preis, T. Yezefski, H. Ouyang, A.A. Suriawinata, A. Silahtaroglu, J.R. Conejo-Garcia, S. Kauppinen, W. Wells, M. Korc, Fluorescence-based codetection with protein markers reveals distinct cellular compartments for altered MicroRNA expression in solid tumours. Clin Cancer Res 16, 4246–4255 (2010)PubMedCentralPubMedCrossRefGoogle Scholar
- 59.L. Qi, J. Bart, L.P. Tan, I. Platteel, T. Sluis, S. Huitema, G. Harms, L. Fu, H. Hollema, A. Berg, Expression of miR-21 and its targets (PTEN, PDCD4, TM1) in flat epithelial atypia of the breast in relation to ductal carcinoma in situ and invasive carcinoma. BMC Cancer 9, 163 (2009)PubMedCentralPubMedCrossRefGoogle Scholar