Feasibility of urinary microRNA detection in breast cancer patients and its potential as an innovative non-invasive biomarker

BMC Cancer

Thalia Erbes²  Marc Hirschfeld⁴  Gerta Rücker¹  Markus Jaeger²  Jasmin Boas²  Severine Iborra²  Sebastian Mayer²  Gerald Gitsch²  Elmar Stickeler³ 

[1] Institute for Medical Biometry and Statistics, University Medical Center Freiburg, Freiburg, Germany

[2] Department of Obstetrics and Gynecology, University Medical Center Freiburg, Hugstetterstr. 55, Freiburg 79106, Germany

[3] German Cancer Consortium (DKTK), Heidelberg, Germany

[4] German Cancer Research Center (DKFZ), Heidelberg, Germany

关键词: Discrimination;    Innovative;    Non-invasive;    Biomarker;    Urine;    microRNA;    Breast cancer;   

Others  :  1161237
DOI  :  10.1186/s12885-015-1190-4

 received in 2014-09-02, accepted in 2015-03-13,  发布年份 2015

【 摘 要 】

Background

Since recent studies revealed the feasibility to detect blood-based microRNAs (miRNAs, miRs) in breast cancer (BC) patients a new field has been opened for circulating miRNAs as potential biomarkers in BC. In this pilot study, we evaluated to our knowledge for the first time whether distinct pattern of urinary miRNAs might be also applicable as innovative biomarkers for BC detection.

Methods

Urinary miRNA expression levels of nine BC-related miRNAs (miR-21, miR-34a, miR-125b, miR-155, miR-195, miR-200b, miR-200c, miR-375, miR-451) from 24 untreated, primary BC patients and 24 healthy controls were quantified by realtime-PCR. The receiver operating characteristic analyses (ROC) and logistic regression were calculated to assess discriminatory accuracy.

Results

Significant differences were found in the expression of four BC-associated miRNAs quantified as median miRNA expression levels. Urinary miR-155 levels were significantly higher in BC patients compared to healthy controls (1.49vs.0.25; p < 0.001). In contrast, compared to healthy controls, BC patients exhibited significantly lower urinary expression levels of miR-21 (2.27vs.5.07; p < 0.001), miR-125b (0.71vs.1.62; p < 0.001), and miR-451 (0.02vs.0.59 p = 0.004), respectively. The ROC including all miRNAs as well as the group of the four significant deregulated miRNAs separated BC patients from healthy controls with a very high (area under the receiver operating characteristic curve [AUC] = 0.932) and high accuracy (AUC = 0.887), respectively.

Conclusions

We were able to demonstrate for the first time the feasibility to detect distinct BC-dependent urinary miRNA profiles. The expression levels of four urinary miRNAs were specifically altered in our cohort of BC patients compared to healthy controls. This distinct pattern offers the possibility for a specific discrimination between healthy women and primary BC patients. This sustains the potential role of urinary miRNAs as non-invasive innovative urine-based biomarkers for BC detection.

【 授权许可】

   
2015 Erbes et al.; licensee BioMed Central.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

【 参考文献 】

• [1]Lewis BP, Burge CB, Bartel DP: Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 2005, 120(1):15-20.
• [2]Sandhu S, Garzon R: Potential applications of microRNAs in cancer diagnosis, prognosis, and treatment. Semin Oncol 2011, 38(6):781-787.
• [3]Thorsen SB, Obad S, Jensen NF, Stenvang J, Kauppinen S: The therapeutic potential of microRNAs in cancer. Cancer J 2012, 18(3):275-284.
• [4]Mulrane L, McGee SF, Gallagher WM, O'Connor DP: miRNA dysregulation in breast cancer. Cancer Res 2013, 73(22):6554-6562.
• [5]Jung M, Schaefer A, Steiner I, Kempkensteffen C, Stephan C, Erbersdobler A, Jung K: Robust microRNA stability in degraded RNA preparations from human tissue and cell samples. Clin Chem 2010, 56(6):998-1006.
• [6]Mattie MD, Benz CC, Bowers J, Sensinger K, Wong L, Scott GK, Fedele V, Ginzinger D, Getts R, Haqq C: Optimized high-throughput microRNA expression profiling provides novel biomarker assessment of clinical prostate and breast cancer biopsies. Molecular cancer 2006, 5:24. BioMed Central Full Text
• [7]Ajit SK: Circulating microRNAs as biomarkers, therapeutic targets, and signaling molecules. Sensors (Basel) 2012, 12(3):3359-3369.
• [8]Turchinovich A, Weiz L, Langheinz A, Burwinkel B: Characterization of extracellular circulating microRNA. Nucleic Acids Res 2011, 39(16):7223-7233.
• [9]Mlcochova H, Hezova R, Stanik M, Slaby O: Urine microRNAs as potential noninvasive biomarkers in urologic cancers. Urol Onco 2014, 32(1):41 e41-49.
• [10]Asaga S, Kuo C, Nguyen T, Terpenning M, Giuliano AE, Hoon DS: Direct serum assay for microRNA-21 concentrations in early and advanced breast cancer. Clin Chem 2011, 57(1):84-91.
• [11]Cortez MA, Bueso-Ramos C, Ferdin J, Lopez-Berestein G, Sood AK, Calin GA: MicroRNAs in body fluids–the mix of hormones and biomarkers. Nat Rev Clin Oncol 2011, 8(8):467-477.
• [12]Heneghan HM, Miller N, Lowery AJ, Sweeney KJ, Newell J, Kerin MJ: Circulating microRNAs as novel minimally invasive biomarkers for breast cancer. Ann Surg 2010, 251(3):499-505.
• [13]Roth C, Rack B, Muller V, Janni W, Pantel K, Schwarzenbach H: Circulating microRNAs as blood-based markers for patients with primary and metastatic breast cancer. Breast Cancer Res 2010, 12(6):R90. BioMed Central Full Text
• [14]Wu X, Somlo G, Yu Y, Palomares MR, Li AX, Zhou W, Chow A, Yen Y, Rossi JJ, Gao H, et al.: De novo sequencing of circulating miRNAs identifies novel markers predicting clinical outcome of locally advanced breast cancer. J Transl Med 2012, 10:42. BioMed Central Full Text
• [15]Chen J, Wang X: MicroRNA-21 in breast cancer: diagnostic and prognostic potential. Clin Trans Oncol 2014, 16(3):225-233.
• [16]Petrovic N, Mandusic V, Stanojevic B, Lukic S, Todorovic L, Roganovic J, Dimitrijevic B: The difference in miR-21 expression levels between invasive and non-invasive breast cancers emphasizes its role in breast cancer invasion. Med Oncol 2014, 31(3):867.
• [17]Bojmar L, Karlsson E, Ellegard S, Olsson H, Bjornsson B, Hallbook O, Larsson M, Stal O, Sandstrom P: The role of microRNA-200 in progression of human colorectal and breast cancer. PloS one 2013, 8(12):e84815.
• [18]Madhavan D, Zucknick M, Wallwiener M, Cuk K, Modugno C, Scharpff M, Schott S, Heil J, Turchinovich A, Yang R, et al.: Circulating miRNAs as surrogate markers for circulating tumor cells and prognostic markers in metastatic breast cancer. Clin Cancer Res 2012, 18(21):5972-5982.
• [19]Heneghan HM, Miller N, Kelly R, Newell J, Kerin MJ: Systemic miRNA-195 differentiates breast cancer from other malignancies and is a potential biomarker for detecting noninvasive and early stage disease. Oncologist 2010, 15(7):673-682.
• [20]Mattiske S, Suetani RJ, Neilsen PM, Callen DF: The oncogenic role of miR-155 in breast cancer. Cancer Epidemiol, Bio Prevent 2012, 21(8):1236-1243.
• [21]Sochor M, Basova P, Pesta M, Dusilkova N, Bartos J, Burda P, Pospisil V, Stopka T: Oncogenic MicroRNAs: miR-155, miR-19a, miR-181b, and miR-24 enable monitoring of early breast cancer in serum. BMC Cancer 2014, 14:448. BioMed Central Full Text
• [22]Zhao FL, Dou YC, Wang XF, Han DC, Lv ZG, Ge SL, Zhang YK: Serum microRNA-195 is down-regulated in breast cancer: a potential marker for the diagnosis of breast cancer. Mol Biol Rep 2014, 41(9):5913-5922.
• [23]Pecot CV, Rupaimoole R, Yang D, Akbani R, Ivan C, Lu C, Wu S, Han HD, Shah MY, Rodriguez-Aguayo C, et al.: Tumour angiogenesis regulation by the miR-200 family. Nat Commun 2013, 4:2427.
• [24]Ng EK, Li R, Shin VY, Jin HC, Leung CP, Ma ES, Pang R, Chua D, Chu KM, Law WL, et al.: Circulating microRNAs as specific biomarkers for breast cancer detection. PloS one 2013, 8(1):e53141.
• [25]Frankel LB, Christoffersen NR, Jacobsen A, Lindow M, Krogh A, Lund AH: Programmed cell death 4 (PDCD4) is an important functional target of the microRNA miR-21 in breast cancer cells. J Biol Chem 2008, 283(2):1026-1033.
• [26]Gong C, Yao Y, Wang Y, Liu B, Wu W, Chen J, Su F, Yao H, Song E: Up-regulation of miR-21 mediates resistance to trastuzumab therapy for breast cancer. J Biol Chem 2011, 286(21):19127-19137.
• [27]Si H, Sun X, Chen Y, Cao Y, Chen S, Wang H, Hu C: Circulating microRNA-92a and microRNA-21 as novel minimally invasive biomarkers for primary breast cancer. J Cancer Res Clin Oncol 2013, 139(2):223-229.
• [28]Yan LX, Huang XF, Shao Q, Huang MY, Deng L, Wu QL, Zeng YX, Shao JY: MicroRNA miR-21 overexpression in human breast cancer is associated with advanced clinical stage, lymph node metastasis and patient poor prognosis. Rna 2008, 14(11):2348-2360.
• [29]Zhu S, Si ML, Wu H, Mo YY: MicroRNA-21 targets the tumor suppressor gene tropomyosin 1 (TPM1). J Biol Chem 2007, 282(19):14328-14336.
• [30]Kastl L, Brown I, Schofield AC: miRNA-34a is associated with docetaxel resistance in human breast cancer cells. Breast Cancer Res Treat 2012, 131(2):445-454.
• [31]Mar-Aguilar F, Mendoza-Ramirez JA, Malagon-Santiago I, Espino-Silva PK, Santuario-Facio SK, Ruiz-Flores P, Rodriguez-Padilla C, Resendez-Perez D: Serum circulating microRNA profiling for identification of potential breast cancer biomarkers. Dis Markers 2013, 34(3):163-169.
• [32]Scott GK, Goga A, Bhaumik D, Berger CE, Sullivan CS, Benz CC: Coordinate suppression of ERBB2 and ERBB3 by enforced expression of micro-RNA miR-125a or miR-125b. J Biol Chem 2007, 282(2):1479-1486.
• [33]Wang H, Tan G, Dong L, Cheng L, Li K, Wang Z, Luo H: Circulating MiR-125b as a marker predicting chemoresistance in breast cancer. PloS one 2012, 7(4):e34210.
• [34]Johansson J, Berg T, Kurzejamska E, Pang MF, Tabor V, Jansson M, Roswall P, Pietras K, Sund M, Religa P, et al.: MiR-155-mediated loss of C/EBPbeta shifts the TGF-beta response from growth inhibition to epithelial-mesenchymal transition, invasion and metastasis in breast cancer. Oncogene 2013, 32(50):5614-5624.
• [35]Sun Y, Cai J, Ma F, Lu P, Huang H, Zhou J: miR-155 mediates suppressive effect of progesterone on TLR3, TLR4-triggered immune response. Immunol Lett 2012, 146(1–2):25-30.
• [36]Li X, Roslan S, Johnstone CN, Wright JA, Bracken CP, Anderson M, Bert AG, Selth LA, Anderson RL, Goodall GJ, et al.: MiR-200 can repress breast cancer metastasis through ZEB1-independent but moesin-dependent pathways. Oncogene 2014, 33(31):4077-4088.
• [37]Gregory PA, Bert AG, Paterson EL, Barry SC, Tsykin A, Farshid G, Vadas MA, Khew-Goodall Y, Goodall GJ: The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1. Nat Cell Biol 2008, 10(5):593-601.
• [38]Bergamaschi A, Katzenellenbogen BS: Tamoxifen downregulation of miR-451 increases 14-3-3zeta and promotes breast cancer cell survival and endocrine resistance. Oncogene 2012, 31(1):39-47.
• [39]Chan M, Liaw CS, Ji SM, Tan HH, Wong CY, Thike AA, Tan PH, Ho GH, Lee AS: Identification of circulating microRNA signatures for breast cancer detection. Clin Cancer Res 2013, 19(16):4477-4487.
• [40]Pfaffl MW, Tichopad A, Prgomet C, Neuvians TP: Determination of stable housekeeping genes, differentially regulated target genes and sample integrity: BestKeeper–Excel-based tool using pair-wise correlations. Biotechnol Lett 2004, 26(6):509-515.
• [41]Brase JC, Wuttig D, Kuner R, Sultmann H: Serum microRNAs as non-invasive biomarkers for cancer. Molecular Cancer 2010, 9:306. BioMed Central Full Text
• [42]Davoren PA, McNeill RE, Lowery AJ, Kerin MJ, Miller N: Identification of suitable endogenous control genes for microRNA gene expression analysis in human breast cancer. BMC Mol Biol 2008, 9:76. BioMed Central Full Text
• [43]Mestdagh P, Van Vlierberghe P, De Weer A, Muth D, Westermann F, Speleman F, Vandesompele J: A novel and universal method for microRNA RT-qPCR data normalization. Genome Biol 2009, 10(6):R64. BioMed Central Full Text
• [44]Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F: Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 2002, 3(7):RESEARCH 0034. BioMed Central Full Text
• [45]Faraoni I, Antonetti FR, Cardone J, Bonmassar E: miR-155 gene: a typical multifunctional microRNA. Biochim Biophys Acta 2009, 1792(6):497-505.
• [46]Christodoulatos GS, Dalamaga M: Micro-RNAs as clinical biomarkers and therapeutic targets in breast cancer: Quo vadis? World J Clin Oncol 2014, 5(2):71-81.
• [47]Iorio MV, Ferracin M, Liu CG, Veronese A, Spizzo R, Sabbioni S, Magri E, Pedriali M, Fabbri M, Campiglio M, et al.: MicroRNA gene expression deregulation in human breast cancer. Cancer Res 2005, 65(16):7065-7070.
• [48]Madhavan D, Cuk K, Burwinkel B, Yang R: Cancer diagnosis and prognosis decoded by blood-based circulating microRNA signatures. Front Genet 2013, 4:116.
• [49]Zhu H, Wu H, Liu X, Evans BR, Medina DJ, Liu CG, Yang JM: Role of MicroRNA miR-27a and miR-451 in the regulation of MDR1/P-glycoprotein expression in human cancer cells. Biochem Pharmacol 2008, 76(5):582-588.
• [50]Weber JA, Baxter DH, Zhang S, Huang DY, Huang KH, Lee MJ, Galas DJ, Wang K: The microRNA spectrum in 12 body fluids. Clin Chem 2010, 56(11):1733-1741.
• [51]Cheng L, Sun X, Scicluna BJ, Coleman BM, Hill AF: Characterization and deep sequencing analysis of exosomal and non-exosomal miRNA in human urine. Kidney Int 2013, 86(2):433-444.
• [52]Mizuta K, Awazu S, Yasuda T, Kishi K: Purification and characterization of three ribonucleases from human kidney: comparison with urine ribonucleases. Arch Biochem Biophy 1990, 281(1):144-151.
• [53]Spencer JD, Schwaderer AL, Dirosario JD, McHugh KM, McGillivary G, Justice SS, Carpenter AR, Baker PB, Harder J, Hains DS: Ribonuclease 7 is a potent antimicrobial peptide within the human urinary tract. Kidney Int 2011, 80(2):174-180.
• [54]Hannafon BN, Ding WQ: Intercellular Communication by Exosome-Derived microRNAs in Cancer. Int J Mol Sci 2013, 14(7):14240-14269.
• [55]Rana S, Malinowska K, Zoller M: Exosomal tumor microRNA modulates premetastatic organ cells. Neoplasia 2013, 15(3):281-295.
• [56]Roma-Rodrigues C, Fernandes AR, Baptista PV: Exosome in Tumour Microenvironment: Overview of the Crosstalk between Normal and Cancer Cells. BioMed Res Int 2014, 2014:179486.