Potentiality of a triple microRNA classifier: miR-193a-3p, miR-23a and miR-338-5p for early detection of colorectal cancer

BMC Cancer

Fung Lin Yong²  Chee Wei Law²  Chee Woon Wang¹ 

[1] Department of Biochemistry, Faculty of Medicine, MAHSA University, 59100 Kuala LumpurMalaysia

[2] Department of Surgery, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia

关键词: MiR-338-5p;    MiR-23a;    MiR-193a-3p;    MicroRNA;    Colorectal cancer;   

Others  :  1079710
DOI  :  10.1186/1471-2407-13-280

 received in 2012-10-06, accepted in 2013-05-14,  发布年份 2013

【 摘 要 】

Background

MicroRNAs (miRNAs) are short, non-coding RNA molecules that act as regulators of gene expression. Circulating blood miRNAs offer great potential as cancer biomarkers. The objective of this study was to correlate the differential expression of miRNAs in tissue and blood in the identification of biomarkers for early detection of colorectal cancer (CRC).

Methods

The study was divided into two phases: (I) Marker discovery by miRNA microarray using paired cancer tissues (n?=?30) and blood samples (CRC, n?=?42; control, n?=?18). (II) Marker validation by stem-loop reverse transcription real time PCR using an independent set of paired cancer tissues (n?=?30) and blood samples (CRC, n?=?70; control, n?=?32). Correlation analysis was determined by Pearson’s test. Logistic regression and receiver operating characteristics curve analyses were applied to obtain diagnostic utility of the miRNAs.

Results

Seven miRNAs (miR-150, miR-193a-3p, miR-23a, miR-23b, miR-338-5p, miR-342-3p and miR-483-3p) have been found to be differentially expressed in both tissue and blood samples. Significant positive correlations were observed in the tissue and blood levels of miR-193a-3p, miR-23a and miR-338-5p. Moreover, increased expressions of these miRNAs were detected in the more advanced stages. MiR-193a-3p, miR-23a and miR-338-5p were demonstrated as a classifier for CRC detection, yielding a receiver operating characteristic curve area of 0.887 (80.0% sensitivity, 84.4% specificity and 83.3% accuracy).

Conclusion

Dysregulations in circulating blood miRNAs are reflective of those in colorectal tissues. The triple miRNA classifier of miR-193a-3p, miR-23a and miR-338-5p appears to be a potential blood biomarker for early detection of CRC.

【 授权许可】

   
2013 Yong et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

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【 参考文献 】

• [1]Worldwide cancer statistics. http://info.cancerresearchuk.org/cancerstats/world/cancer-worldwide-the-global-picture webcite
• [2]Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D: Global cancer statistics. CA Cancer J Clin 2011, 61:69-90.
• [3]Bray F, Jemal A, Grey N, Ferlay J, Forman D: Global cancer transitions according to the Human Development Index (2008–2030): a population-based study. Lancet Oncol 2012, 13(8):790-801.
• [4]American Cancer Society: Cancer Facts & Figures 2012. Atlanta: American Cancer Society; 2012.
• [5]Zainal Ariffin O, Nor Saleha IT: National Cancer Registry Report 2007. Malaysia: Ministry of Healthy; 2011.
• [6]McFarland EG, Levin B, Lieberman DA, Pickhardt PJ, Johnson CD, Glick SN, Brooks D, Smith RA: Revised colorectal screening guidelines: joint effort of the American Cancer Society, U.S. Multisociety Task Force on Colorectal Cancer, and American College of Radiology. Radiology 2008, 248:717-720.
• [7]Tests to detect colorectal cancer and polyps. . http://www.cancer.gov/cancertopics/factsheet/detection/colorectal-screening webcite
• [8]Neri E, Faggioni L, Cerri F, Turini F, Angeli S, Cini L, Perrone F, Paolicchi F, Bartolozzi C: CT colonography versus double-contrast barium enema for screening of colorectal cancer: comparison of radiation burden. Abdom Imaging 2010, 35:596-601.
• [9]Choong MK, Tsafnat G: Genetic and epigenetic biomarkers of colorectal cancer. Clin Gastroenterol Hepatol 2012, 10:9-15.
• [10]Khalid-de Bakker CA, Jonkers DM, Sanduleanu S, de Bruïne AP, Meijer GA, Janssen JB, van Engeland M, Stockbrügger RW, Masclee AA: Test performance of immunologic fecal occult blood testing and sigmoidoscopy compared with primary colonoscopy screening for colorectal advanced adenomas. Cancer Prev Res 2011, 4(10):1563-1571.
• [11]Imperiale TF, Ransohoff DF, Itzkowitz SH, Turnbull BA, Ross ME: Fecal DNA versus fecal occult blood for colorectal-cancer screening in an average risk population. N Engl J Med 2004, 351:2704-2714.
• [12]Zou H, Taylor WR, Harrington JJ, Hussain FTN, Cao X, Loprinzi CL, Levine TR, Rex DK, Ahnen D, Knigge KL, et al.: High detection rates of colorectal neoplasia by stool DNA testing with a novel digital melt curve assay. Gastroenterology 2009, 136:459-470.
• [13]U.S. Preventive Services Task Force: Screening for colorectal cancer: U.S. Preventive Services Task Force recommendation statement. Ann Intern Med 2008, 149(9):627-637.
• [14]Ahlquist DA, Sargent DJ, Loprinzi CL, Levin TR, Rex DK, Ahnen DJ, Knigge K, Lance MP, Burgart LJ, Hamilton SR, et al.: Stool DNA and occult blood testing for screen detection of colorectal neoplasia. Ann Intern Med 2008, 149(7):441-450.
• [15]Bartel DP: MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 2004, 116:281-297.
• [16]Lee YS, Dutta A: MicroRNAs in cancer. Annu Rev Pathol 2009, 4:199-227.
• [17]Li L-C, Okino ST, Zhao H, Pookot D, Place RF, Urakami S, Enokida H, Dahiya R: Small dsRNAs induce transcriptional activation in human cells. Proc Natl Acad Sci USA 2006, 103(46):17337-17342.
• [18]Lee RC, Feinbaum RL, Ambros V: The C.elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14. Cell 1993, 75:843-854.
• [19]Kozamara A, Griffiths-Jones S: miRBase: integrating microRNA annotation and deep-sequencing data. Nucleic Acids Res 2011, 39:D152-D157.
• [20]Esquela-Kerscher A, Slack FJ: Oncomirs - microRNAs with a role in cancer. Nat Rev Cancer 2006, 6:259-269.
• [21]Bueno MJ, Malumbres M: MicroRNA and the cell cycle. Biochim Biophys Acta 2011, 1812(5):592-601.
• [22]Henegan HM, Miller N, Kerin MJ: MiRNAs as biomarkers and therapeutic targets in cancer. Curr Opin Pharmacol 2010, 10:543-550.
• [23]Lima RT, Busacca S, Almeida GM, Gaudino G, Fennell DA, Vasconcelos MH: MicroRNA regulation of core apoptosis pathways in cancer. Eur J Cancer 2011, 47:163-174.
• [24]Calin GA, Sevignani C, Dumitru CD, Hyslop T, Noch E, Yendamuri S, Shimizu M, Rattan S, Bullrich F, Negrini M, et al.: Human microRNA genes are frequently located at fragile sites and genomic regions involved in cancers. Proc Natl Acad Sci USA 2004, 101(9):2999-3004.
• [25]Koturbash I, Zemp FJ, Pogribny I, Kovalchuk O: Small molecules with big effects: the role of the microRNAome in cancer and carcinogenesis. Mutat Res 2011, 722:94-105.
• [26]Babashah S, Soleimani M: The oncogenic and tumour suppresive roles of microRNAs in cancer and apoptosis. Eur J Cancer 2011, 47:1127-1137.
• [27]Groce CM: Causes and consequences of microRNA dysregulation in cancer. Nat Rev Genet 2009, 10:704-714.
• [28]Liu M, Chen H: The role of microRNAs in colorectal cancer. J Genet Genomics 2010, 37:347-358.
• [29]Volinia S, Calin GA, Liu CG, Ambs S, Cimmino A, Petrocca F, Visone R, Lorio M, Roldo C, Ferracin M, et al.: A microRNA expression signature of human solid tumors defines cancer gene targets. Proc Natl Acad Sci USA 2006, 103(7):2257-2261.
• [30]Yu DC, Li QC, Ding XW, Ding YT: Circulating microRNAs: potential biomarkers for cancer. Int J Mol Sci 2011, 12:2055-2063.
• [31]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.
• [32]Link A, Balaguer F, Shen Y, Nagasaka T, Lozano JJ, Boland CR, Goel A: Fecal microRNAs as novel biomarkers for colon cancer screening. Cancer Epidemiol Biomarkers Prev 2010, 19(7):1766-1774.
• [33]Valadi H, Ekstrom K, Bossios A, Sjostrand M, Lee JJ, Lotvall JO: Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol 2007, 9:654-659.
• [34]Hunter MP, Ismail N, Zhang X, Aguda BD, Lee EJ, Yu L, Xiao T, Schafer J, Ting Lee M-L, Schmittgen TD, et al.: Detection of microRNA expression in human peripheral blood microvesicles. PLoS One 2008, 3(11):e3694.
• [35]Di Stefano V, Zaccagnini G, Capogrossi MC, Martelli F: microRNAs as peripheral blood biomarkers of cardiovascular disease. Vascul Pharmacol 2011, 55:111-118.
• [36]Zhang J, Zhao H, Gao Y, Zhang W: Secretory miRNAs as novel cancer biomarkers. Biochim Biophys Acta 2012, 1826(1):32-43.
• [37]Zampetaki A, Kiechl S, Drozdov I, Willeit P, Mayr U, Prokopi M, Mayr A, Weger S, Oberhollenzer F, Bonora E, et al.: Plasma microRNA profiling reveals loss of endothelial miR-126 and other microRNAs in type 2 diabetes. Circ Res 2010, 107:810-817.
• [38]Schrauder MG, Strick R, Schulz-Wendtland R, Strissel PL, Kahmann L, Loehberg CR, Lux MP, Jud SM, Hartmann A, Hein A, et al.: Circulating micro-RNAs as potential blood-based markers for early stage breast cancer detection. PLoS One 2012, 7(1):e29770.
• [39]Heneghan HM, Miller N, Lowery AJ, Sweeney KJ, Newell J: Circulating microRNAs as novel minimally invasive biomarkers for breast cancer. Ann Surg 2010, 251:499-505.
• [40]Kosaka N, Iguchi H, Ochiya T: Circulating microRNA in body fluid: a new potential biomarker for cancer diagnosis and prognosis. Cancer Sci 2010, 101(10):2087-2092.
• [41]Turchinovich A, Weiz L, Langheinz A, Burwinkel B: Characterization of extracellular miRNA. Nucleic Acids Res 2011, 39(16):7223-7233.
• [42]Luo X, Burwinkel B, Tao S, Brenner H: MicroRNA signatures: novel biomarker for colorectal cancer? Cancer Epidemiol Biomarkers Prev 2011, 20(7):1272-1286.
• [43]Schroeder A, Mueller O, Stocker S, Salowsky R, Leiber M, Gassmann M, Lightfoot S, Menzel W, Granzow M, Ragg T: The RIN: an RNA integrity number for assigning integrity values to RNA measurements. BMC Mol Biol 2006, 7:3. BioMed Central Full Text
• [44]Irizarry RA, Hobbs B, Collin F, Beazer-Barclay YD, Antonellis KJ, Scherf U, Speed TP: Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics 2003, 4:249-264.
• [45]Jordan JA, Durso MB: Real-time polymerase chain reaction for detecting bacterial DNA directly from blood of neonates being evaluated for Sepsis. J Mol Diagn 2005, 7(5):575-581.
• [46]Caraguel CGB, Stryhn H, Gagne N, Dohoo IR, Hammell KL: Selection of a cutoff value for real-time polymerase chain reaction results to fit a diagnostic purpose: analytical and epidemiologic approaches. J Vet Diagn Invest 2011, 23:2-15.
• [47]Yuan JS, Reed A, Chen F, Stewart CN Jr: Statistical analysis of real-time PCR data. BMC Bioinformatics 2006, 7:85. BioMed Central Full Text
• [48]Obuchowski NA: ROC analysis. AJR Am J Roentgenol 2005, 184:364-372.
• [49]Piepoli A, Tavano F, Copetti M, Mazza T, Palumbo O, Panza A, di Mola FF, Pazienza V, Mazzoccoli G, Biscaglia G, et al.: Mirna expression profiles identify drivers in colorectal and pancreatic cancers. PLoS One 2012, 7(3):e33663.
• [50]Hamfjord J, Stangeland AM, Hughes T, Skrede ML, Tveit KM, Ikdahl T, Kure EH: Differential expression of miRNAs in colorectal cancer: comparison of paired tumor tissue and adjacent normal mucosa using high-throughput sequencing. PLoS One 2012, 7(4):e34150.
• [51]Ahmed FE, Amed NC, Vos PW, Bonnerup C, Atkins JN, Casey M, Nuovo GJ, Naziri W, WIley JE, Allison RR: Diagnostic microRNA markers to screen for sporadic human colon cancer in blood. Cancer Genomics Proteomics 2012, 9(4):179-192.
• [52]Ng EK, Chong WW, Jin H, Lam EK, Shin VY, Yu J: Differential expression of microRNAs in plasma of patients with colorectal cancer: a potential marker for colorectal cancer screening. Gut 2009, 58:1375-1381.
• [53]Benjamin H, Lebanony D, Rosenwald S, Cohen L, Gibori H, Barabash N, Ashkenazi K, Goren E, Meiri E, Morgenstern S, et al.: A diagnostic assay based on microRNA expression accurately identifies malignant pleural mesothelioma. J Mol Diagn 2010, 12(6):771-779.
• [54]Baumhoer D, Zillmer S, Unger K, Rosemann M, Atkinson MJ, Irmler M, Beckers J, Siggelkow H, Von Luettichau I, Jundt G, et al.: MicroRNA profiling with correlation to gene expression revealed the oncogenic miR-17-92 cluster to be up-regulated in osteosarcoma. Cancer Genet 2012, 205:212-219.
• [55]Kozaki K, Imoto I, Mogi S, Omura K, Inazawa J: Exploration of tumor-suppressive microRNAs silenced by DNA hypermethylation in oral cancer. Cancer Res 2008, 68(7):2094-2105.
• [56]Jahid S, Sun J, Edwards RA, Dizon D, Panarelli NC, Milsom JW, Sikandar SS, Gumus ZH, Lipkin SM: miR-23a promotes the transition from indolent to invasive colorectal cancer. Cancer Discov 2012, 2(6):540-553.
• [57]Chen X, Ba Y, Ma L, Cai X, Yin Y, Wang K, Guo J, Zhang Y, Chen J, Guo X, et al.: Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases. Cell Res 2008, 18:997-1006.
• [58]Zhu LH, Liu T, Tang H, Tian RQ, Su C, Liu M, Li X: MicroRNA-23a promotes the growth of gastric adenocarcinoma cell line MGC803 and downregulates interleukin-6 receptor. FEBS J 2010, 277:3726-3734.
• [59]Chhabra R, Dubey R, Saini N: Cooperative and individualistic functions of the microRNAs in the miR23a?~?27a?~?24-2 cluster and its implication in human diseases. Mol Cancer 2010, 9:232. BioMed Central Full Text
• [60]Willert K, Jones KA: Wnt signaling: is the party in the nucleus? Genes Dev 2006, 20:1394-1404.
• [61]Rogler CE, LeVoci L, Ader T, Massimi A, Tchaikovskaya T, Norel R, Rogler LE: MicroRNA-23b cluster microRNAs regulate transforming growth factor-beta/bone morphogenetic protein signaling and liver stem cell differentiation by targeting smads. Hepatology 2009, 50:575-584.
• [62]Gao P, Tchernyshyov I, Chang TC, Lee YS, Kita K, Ochi T, Zeller K, De Marzo AM, Van Eyk JE, Mendell JT, et al.: c-Myc suppression of miR-23a/b enhances mitochondrial glutaminase and glutamine metabolism. Nature 2009, 458(7239):762-765.
• [63]Saumet A, Vetter G, Bouttier M, Portales-Casamar E, Wasserman WW, Maurin T, Mari B, Barbry P, Vallar L, Friederich E, et al.: Transcriptional repression of microRNA genes by PML-RARA increases expression of key cancer proteins in acute promyelocytic leukemia. Blood 2009, 113(2):412-421.
• [64]Schetter AJ, Leung SY, Sohn JJ, Zanetti KA, Bowman ED, Yanaihara N, Yuen ST, Chan TL, Kwong DLW, Au GKH, et al.: MicroRNA expression profiles associated with prognosis and therapeutic outcome in colon adenocarcinoma. JAMA 2008, 299(4):425-436.
• [65]Budhu A, Jia HL, Forgues M, Liu CG, Goldstein D, Lam A, Zanetti KA, Ye QH, Qin LX, Croce CM, et al.: Identification of metastasis-related microRNAs in hepatocellular carcinoma. Hepatology 2008, 47(3):897-907.
• [66]Wong TS, Liu XB, Wong BYH, Ng WM, Yuen APW, Wei WI: Mature miR-184 as potential oncogenic microRNA of squamous cell carcinoma of tongue. Clin Cancer Res 2008, 14(9):2588-2592.
• [67]Ju JA, Huang YC, Lan SH, Wang TH, Lin PC, Lee JC, Niu KC, Tian YF, Liu HS: Identification of colorectal cancer recurrence-related microRNAs. GMBHS 2012, 4:19-20.