Tumor Biology

, Volume 36, Issue 11, pp 8845–8852 | Cite as

Identification of miR-199a-5p in serum as noninvasive biomarkers for detecting and monitoring osteosarcoma

Research Article


Emerging evidence has suggested that circulating microRNAs (miRNAs) in serum/plasma can serve as noninvasive biomarkers for cancer detection; however, little is known about circulating miRNA profiles in osteosarcoma, a primary malignant bone tumor with high morbidity in young adults and adolescents. The objective of this study was to investigate whether circulating miRNAs in serum could be a useful biomarker for detecting osteosarcoma and monitoring tumor dynamics. Serum samples were obtained from 60 patients before surgery, 28 patients after 1 month of surgery, and 60 healthy individuals. The study was divided into three steps: (1) initial screening of the profiles of circulating miRNAs in pooled serum samples from both healthy controls and pre- and postoperative osteosarcoma patients using a TaqMan low-density qPCR array (TLDA); (2) evaluation of miRNA concentration in individual serum samples from 60 preoperative osteosarcoma patients and 60 healthy controls by a quantitative RT-PCR assay; and (3) evaluation of miRNA concentration in paired serum samples from 28 pre- and postoperative osteosarcoma patients by a quantitative RT-PCR assay. The initial analysis showed that concentrations of serum miRNAs were significantly altered between preoperative osteosarcoma patients and healthy controls and between pre- and postoperative osteosarcoma patients. The quantitative RT-PCR assay showed that serum miR-199a-5p concentrations were significantly higher in osteosarcoma patients than in controls. The value of the area under the ROC curve was 0.8606. Serum levels of miR-199a-5p were significantly lower in post- than preoperative samples. The results indicated the potential of circulating miRNAs as novel noninvasive biomarkers for detecting and monitoring osteosarcoma.


MicroRNA Serum microRNA Osteosarcoma miR-199a-5p 



This work was supported by grants from the National Natural Science Foundation of China (no. 81102039), China Postdoctoral Science Foundation (no. 2012 M512178), and Special Fund of Jiangsu Province for the Clinical Center of Orthopedics (no. BL2012002). The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Conflicts of interest


Supplementary material

13277_2015_3421_MOESM1_ESM.doc (288 kb)
ESM 1 (DOC 288 kb)


  1. 1.
    Marina N, Gebhardt M, Teot L, Gorlick R. Biology and therapeutic advances for pediatric osteosarcoma. Oncologist. 2004;9:422–41.CrossRefPubMedGoogle Scholar
  2. 2.
    Bielack SS, Marina N, Ferrari S, Helman LJ, Smeland S, et al. Osteosarcoma: the same old drugs or more? J Clin Oncol. 2008;26:3102–3. author reply 3104–3105.CrossRefPubMedGoogle Scholar
  3. 3.
    Longhi A, Errani C, De Paolis M, Mercuri M, Bacci G. Primary bone osteosarcoma in the pediatric age: state of the art. Cancer Treat Rev. 2006;32:423–36.CrossRefPubMedGoogle Scholar
  4. 4.
    Calin GA, Croce CM. MicroRNA signatures in human cancers. Nat Rev Cancer. 2006;6:857–66.CrossRefPubMedGoogle Scholar
  5. 5.
    Esquela-Kerscher A, Slack FJ. Oncomirs—microRNAs with a role in cancer. Nat Rev Cancer. 2006;6:259–69.CrossRefPubMedGoogle Scholar
  6. 6.
    Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004;116:281–97.CrossRefPubMedGoogle Scholar
  7. 7.
    He L, Hannon GJ. MicroRNAs: small RNAs with a big role in gene regulation. Nat Rev Genet. 2004;5:522–31.CrossRefPubMedGoogle Scholar
  8. 8.
    Mitchell PS, Parkin RK, Kroh EM, Fritz BR, Wyman SK, et al. Circulating microRNAs as stable blood-based markers for cancer detection. Proc Natl Acad Sci U S A. 2008;105:10513–8.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Chen X, Ba Y, Ma L, Cai X, Yin Y, 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.CrossRefPubMedGoogle Scholar
  10. 10.
    Lawrie CH, Gal S, Dunlop HM, Pushkaran B, Liggins AP, et al. Detection of elevated levels of tumour-associated microRNAs in serum of patients with diffuse large B-cell lymphoma. Br J Haematol. 2008;141:672–5.CrossRefPubMedGoogle Scholar
  11. 11.
    Ouyang L, Liu P, Yang S, Ye S, Xu W, et al. A three-plasma miRNA signature serves as novel biomarkers for osteosarcoma. Med Oncol. 2013;30:340.CrossRefPubMedGoogle Scholar
  12. 12.
    Shen Q, Cicinnati VR, Zhang X, Iacob S, Weber F, et al. Role of microRNA-199a-5p and discoidin domain receptor 1 in human hepatocellular carcinoma invasion. Mol Cancer. 2010;9:227.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Iorio MV, Visone R, Di Leva G, Donati V, Petrocca F, et al. MicroRNA signatures in human ovarian cancer. Cancer Res. 2007;67:8699–707.CrossRefPubMedGoogle Scholar
  14. 14.
    Cheng W, Liu T, Wan X, Gao Y, Wang H. MicroRNA-199a targets CD44 to suppress the tumorigenicity and multidrug resistance of ovarian cancer-initiating cells. FEBS J. 2012;279:2047–59.CrossRefPubMedGoogle Scholar
  15. 15.
    Duan Z, Choy E, Harmon D, Liu X, Susa M, et al. MicroRNA-199a-3p is downregulated in human osteosarcoma and regulates cell proliferation and migration. Mol Cancer Ther. 2011;10:1337–45.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Eis PS, Tam W, Sun L, Chadburn A, Li Z, et al. Accumulation of miR-155 and BIC RNA in human B cell lymphomas. Proc Natl Acad Sci U S A. 2005;102:3627–32.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Fang C, Zhu DX, Dong HJ, Zhou ZJ, Wang YH, et al. Serum microRNAs are promising novel biomarkers for diffuse large B cell lymphoma. Ann Hematol. 2012;91:553–9.CrossRefPubMedGoogle Scholar
  18. 18.
    Cho WC, Chow AS, Au JS. Restoration of tumour suppressor hsa-miR-145 inhibits cancer cell growth in lung adenocarcinoma patients with epidermal growth factor receptor mutation. Eur J Cancer. 2009;45:2197–206.CrossRefPubMedGoogle Scholar
  19. 19.
    Chen X, Hu Z, Wang W, Ba Y, Ma L, et al. Identification of ten serum microRNAs from a genome-wide serum microRNA expression profile as novel noninvasive biomarkers for nonsmall cell lung cancer diagnosis. Int J Cancer. 2012;130:1620–8.CrossRefPubMedGoogle Scholar
  20. 20.
    Ohshima K, Inoue K, Fujiwara A, Hatakeyama K, Kanto K, et al. Let-7 microRNA family is selectively secreted into the extracellular environment via exosomes in a metastatic gastric cancer cell line. PLoS One. 2010;5:e13247.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  1. 1.Department of Orthopedics, Jinling Hospital, School of MedicineNanjing UniversityNanjingChina
  2. 2.State Key Laboratory of Pharmaceutical Biotechnology, School of Life SciencesNanjing UniversityNanjingChina

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