The Upregulation of hsa-mir-181b-1 and Downregulation of Its Target CYLD in the Late-Stage of Tumor Progression of Breast Cancer


Some microRNAs are usually dysregulated in the cancers and influencing tumor behavior and progression. Hsa-miR-181b-1 and its target CYLD are involved in regulating the inflammatory pathways. This study aimed to investigate the expression levels of hsa-mir-181b-1 and CYLD in a cohort of breast tumor tissues and normal adjacent tissues to assess their association with breast cancer stages. A total number of 60 breast samples including cancerous and normal adjacent tissue specimens were collected. After pathological study, the expression of hsa-mir-181b-1 and CYLD were measured by qRT-PCR method. The hsa-mir-181b-1 expression level was significantly increased in breast tumor tissues compared to the controls. This increase was associated with the disease progression. Conversely, CYLD expression level was decreased in tumor samples compared to normal samples, significantly. ROC curve data added other prestigious information of hsa-mir-181b-1 and CYLD by defining cancer and healthy tissues with high specificity and sensitivity at a proposed cutoff point. Also, bioinformatic enrichment for the possible targets of mature sequence of “hsa-mir-181b-5p” was performed. Computational analysis showed the five most significant pathways including metabolic, cancer, calcium signaling, PI3K–Akt signaling and focal adhesion pathways which may be influenced by hsa-mir-181b-1. Thus, we suggested hsa-mir-181b-1 and CYLD might be involved in the pathogenesis of breast cancer and could be considered as two biomarkers for prediction, prognosis and diagnosis of the stages of the breast cancer.

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  1. 1.

    Iliopoulos D, Jaeger SA, Hirsch HA, Bulyk ML, Struhl K. STAT3 activation of miR-21 and miR-181b-1 via PTEN and CYLD are part of the epigenetic switch linking inflammation to cancer. Mol Cell. 2010;39(4):493–506.

    CAS  Article  Google Scholar 

  2. 2.

    Grivennikov SI, Greten FR, Karin M. Immunity, inflammation, and cancer. Cell. 2010;140(6):883–99.

    CAS  Article  Google Scholar 

  3. 3.

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

    CAS  Article  Google Scholar 

  4. 4.

    Liu J, Shi W, Wu C, Ju J, Jiang J. MiR-181b as a key regulator of the oncogenic process and its clinical implications in cancer. Biomed Rep. 2014;2(1):7–11.

    CAS  Article  Google Scholar 

  5. 5.

    Yan LX, Huang XF, Shao Q, Huang MY, Deng L, Wu QL, et al. MicroRNA miR-21 overexpression in human breast cancer is associated with advanced clinical stage, lymph node metastasis and patient poor prognosis. RNA. 2008;14:2348–60.

    CAS  Article  Google Scholar 

  6. 6.

    Andorfer CA, Necela BM, Thompson EA, Perez EA. MicroRNA signatures: clinical biomarkers for the diagnosis and treatment of breast cancer. Trends Mol Med. 2011;17(6):313–9.

    CAS  Article  Google Scholar 

  7. 7.

    Rouigari M, Dehbashi M, Tabatabaeian H, Ghaedi K, Mohammadynejad P, Azadeh M. Evaluation of the expression level and hormone receptor association of miR-126 in breast cancer. Indian J Clin Biochem. 2018.

    Article  PubMed  Google Scholar 

  8. 8.

    Li D, Jian W, Wei C, Song H, Gu Y, Luo Y, et al. Down-regulation of miR-181b promotes apoptosis by targeting CYLD in thyroid papillary cancer. Int J Clin Exp Pathol. 2014;7(11):7672.

    PubMed  PubMed Central  Google Scholar 

  9. 9.

    Shostak K, Chariot A. NF-κB, stem cells and breast cancer: the links get stronger. Breast Cancer Res. 2011;13(4):214.

    Article  Google Scholar 

  10. 10.

    Shi L, Cheng Z, Zhang J, Li R, Zhao P, Fu Z, et al. Hsa-mir-181a and hsa-mir-181b function as tumor suppressors in human glioma cells. Brain Res. 2008;1236:185–93.

    CAS  Article  Google Scholar 

  11. 11.

    Zanette DL, Rivadavia F, Molfetta GA, Barbuzano FG, Proto-Siqueira R, Silva WA Jr, et al. miRNA expression profiles in chronic lymphocytic and acute lymphocytic leukemia. Braz J Med Biol Res. 2007;40(11):1435–40.

    CAS  Article  Google Scholar 

  12. 12.

    Ren Y, Gao J, Liu JQ, Wang XW, Gu JJ, Huang HJ, et al. Differential signature of fecal microRNAs in patients with pancreatic cancer. Mol Med Rep. 2012;6(1):201–9.

    CAS  PubMed  Google Scholar 

  13. 13.

    Nurul-Syakima Α, Yoke-Kqueen C, Sabariah A, Shiran M, Singh A, et al. Differential microRNA expression and identification of putative miRNA targets and pathways in head and neck cancers. Int J Mol Med. 2011;28(3):327–36.

    PubMed  Google Scholar 

  14. 14.

    Ratert N, Meyer HA, Jung M, Mollenkopf HJ, Wagner I, Miller K, et al. Reference miRNAs for miRNAome analysis of urothelial carcinomas. PLoS ONE. 2012;7(6):e39309.

    CAS  Article  Google Scholar 

  15. 15.

    Li X, Zhang Y, Zhang H, Liu X, Gong T, Li M, et al. MiRNA-223 promotes gastric cancer invasion and metastasis by targeting tumor suppressor EPB41L3. Mol Cancer Res. 2011;9(7):824–33.

    CAS  Article  Google Scholar 

  16. 16.

    Schaefer A, Jung M, Mollenkopf HJ, Wagner I, Stephan C, Jentzmik F, et al. Diagnostic and prognostic implications of microRNA profiling in prostate carcinoma. Int J Cancer. 2010;126(5):1166–76.

    CAS  PubMed  Google Scholar 

  17. 17.

    Sun X, Icli B, Wara AK, Belkin N, He S, Kobzik L, et al. MicroRNA-181b regulates NF-κB-mediated vascular inflammation. J Clin Investig. 2012;122(6):1973–90.

    CAS  PubMed  Google Scholar 

  18. 18.

    Wolff AC, Hammond ME, Schwartz JN, Hagerty KL, Allred DC, Cote RJ, et al. American Society of Clinical Oncology/College of American Pathologists guideline recommendations for human epidermal growth factor receptor 2 testing in breast cancer. Arch Pathol Lab Med. 2007;131:18–43.

    CAS  PubMed  Google Scholar 

  19. 19.

    Ciafre SA, Galardi S, Mangiola A, Ferracin M, Liu CG, Sabatino G, et al. Extensive modulation of a set of microRNAs in primary glioblastoma. Biochem Biophys Res Commun. 2005;334(4):1351–8.

    CAS  Article  Google Scholar 

  20. 20.

    Xi Y, Formentini A, Chien M, Weir DB, Russo JJ, Ju J, et al. Prognostic values of microRNAs in colorectal cancer. Biomark Insights. 2006;2:113–21.

    PubMed  Google Scholar 

  21. 21.

    Jiang J, Zheng X, Xu X, Zhou Q, Yan H, Zhang X, et al. Prognostic significance of miR-181b and miR-21 in gastric cancer patients treated with S-1/oxaliplatin or doxifluridine/oxaliplatin. PLoS ONE. 2011;6(8):e23271.

    CAS  Article  Google Scholar 

  22. 22.

    Wu L, Lin Q, Shi J, Lin X. Evaluation of miR-182/miR-100 ratio for diagnosis and survival prediction in bladder cancer. Arch Iran Med. 2016;19(9):645.

    PubMed  Google Scholar 

  23. 23.

    Conti A, Aguennouz M, La Torre D, Tomasello C, Cardali S, Angileri FF, et al. MiR-21 and 221 upregulation and miR-181b downregulation in human grade II–IV astrocytic tumors. J Neurooncol. 2009;93(3):325–32.

    CAS  Article  Google Scholar 

  24. 24.

    Xu DD, Zhou PJ, Wang Y, Zhang L, Fu WY, Ruan BB, et al. Reciprocal activation between STAT3 and miR-181b regulates the proliferation of esophageal cancer stem-like cells via the CYLD pathway. Mol Cancer. 2016;15(1):40.

    Article  Google Scholar 

  25. 25.

    Ghadami E, Nikbakhsh N, Fattahi S, Kosari-Monfared M, Ranaee M, Taheri H, et al. Epigenetic alterations of CYLD promoter modulate its expression in gastric adenocarcinoma: a footprint of infections. J Cell Physiol. 2019;234(4):4115–24.

    CAS  Article  Google Scholar 

  26. 26.

    Kinoshita H, Okabe H, Beppu T, Chikamoto A, Hayashi H, Imai K, et al. CYLD downregulation is correlated with tumor development in patients with hepatocellular carcinoma. Mol Clin Oncol. 2013;1(2):309–14.

    Article  Google Scholar 

  27. 27.

    Hutti JE, Shen RR, Abbott DW, Zhou AY, Sprott KM, Asara JM, et al. Phosphorylation of the tumor suppressor CYLD by the breast cancer oncogene IKKepsilon promotes cell transformation. Mol Cell. 2009;34(4):461–72.

    CAS  Article  Google Scholar 

  28. 28.

    Volinia S, Calin GA, Liu CG, Ambs S, Cimmino A, Petrocca F, et al. A microRNA expression signature of human solid tumors defines cancer gene targets. Proc Natl Acad Sci USA. 2006;103(7):2257–61.

    CAS  Article  Google Scholar 

  29. 29.

    Bloomston M, Frankel WL, Petrocca F, Volinia S, Alder H, Hagan JP, et al. MicroRNA expression patterns to differentiate pancreatic adenocarcinoma from normal pancreas and chronic pancreatitis. JAMA. 2007;297(17):1901–8.

    CAS  Article  Google Scholar 

  30. 30.

    Ji J, Yamashita T, Budhu A, Forgues M, Jia HL, Li C, et al. Identification of microRNA-181 by genome-wide screening as a critical player in EpCAM-positive hepatic cancer stem cells. Hepatology. 2009;50(2):472–80.

    CAS  Article  Google Scholar 

  31. 31.

    Wang F, Sun JY, Zhu YH, Liu NT, Wu YF, Yu F. MicroRNA-181 inhibits glioma cell proliferation by targeting cyclin B1. Mol Med Rep. 2014;10(4):2160–4.

    CAS  Article  PubMed  Google Scholar 

  32. 32.

    Huang P, Ye B, Yang Y, Shi J, Zhao H. MicroRNA-181 functions as a tumor suppressor in non-small cell lung cancer (NSCLC) by targeting Bcl-2. Tumor Biol. 2015;36(5):3381–7.

    CAS  Article  Google Scholar 

  33. 33.

    Tong SJ, Liu J, Wang X, Qu LX. MicroRNA-181 promotes prostate cancer cell proliferation by regulating DAX-1 expression. Exp Ther Med. 2014;8(4):1296–300.

    CAS  Article  Google Scholar 

  34. 34.

    Sun Y, Yu S, Liu Y, Wang F, Liu Y, Xiao H. Expression of miRNAs in papillary thyroid carcinomas is associated with BRAF mutation and clinicopathological features in Chinese patients. Int J Endocrinol. 2013;2013.

  35. 35.

    Salaun B, Yamamoto T, Badran B, Tsunetsugu-Yokota Y, Roux A, Baitsch L, et al. Differentiation associated regulation of microRNA expression in vivo in human CD8+ T cell subsets. J Transl Med. 2011;9(1):44.

    CAS  Article  Google Scholar 

  36. 36.

    Pallante P, Visone R, Ferracin M, Ferraro A, Berlingieri MT, Troncone G, et al. MicroRNA deregulation in human thyroid papillary carcinomas. Endocr Relat Cancer. 2006;13(2):497–508.

    CAS  Article  Google Scholar 

  37. 37.

    Peng J, Thakur A, Zhang S, Dong Y, Wang X, Yuan R, et al. Expressions of miR-181a and miR-20a in RPMI8226 cell line and their potential as biomarkers for multiple myeloma. Tumor Biol. 2015;36(11):8545–52.

    CAS  Article  Google Scholar 

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The authors would like to thank for the Cancer Institute of Imam Khomeini Hospital, Tehran University of Medical Sciences, Tehran, Iran for providing the clinical samples and also for the graduate offices of Tehran University of Medical Sciences and Isfahan University of Medical Sciences for their financial supports (Grant No. 192046).

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Correspondence to Mazdak Ganjalikhani-Hakemi.

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Andalib, A., Rashed, S., Dehbashi, M. et al. The Upregulation of hsa-mir-181b-1 and Downregulation of Its Target CYLD in the Late-Stage of Tumor Progression of Breast Cancer. Ind J Clin Biochem 35, 312–321 (2020).

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  • Breast cancer
  • hsa-mir-181b-1
  • CYLD
  • Expression level
  • Inflammation