Exosomal miR-1246 in serum as a potential biomarker for early diagnosis of gastric cancer

  • Yuntao  Shi
  • Zhonghong  Wang
  • Xiaojuan  Zhu
  • Ling  Chen
  • Yilan  Ma
  • Jiayan  Wang
  • Xiaozhong  YangEmail author
  • Zheng  LiuEmail author
Original Article



Gastric cancer (GC) patients are usually diagnosed in advanced stages which results in high mortality. This study aimed to identify novel circulating miRNAs as biomarkers for the early detection of GC.


Candidate miRNA was identified after integrated analysis of two Gene Expression Omnibus (GEO) datasets and clinical serum samples. Exosomes extracted were verified using transmission electron microscopy (TEM) and western blot. The expressions of miRNAs were tested through qRT-PCR. Receiver operating characteristic curve (ROC) analysis was used to explore the diagnostic utility of miRNAs. RNA pull-down assay was used to find RNA binding proteins (RBPs) which transport candidate miRNA into exosomes. Bioinformatics analysis of candidate miRNA was conducted using DAVID and Cytoscape.


After integrated analysis of two GEO datasets, six circulating miRNAs were found to be consistently upregulated in GC patients. Then, qRT-PCR demonstrated that serum miR-1246 was the one with the largest fold change. Studies in vitro revealed that elevated serum miR-1246 was tumor-derived by being packaged into exosomes with the help of ELAVL1. Thereafter, we discovered that exosomal miR-1246 expressions in serum could differentiate GC patients with TNM stage I from healthy controls (HCs) and patients with benign diseases (BDs) with area under the curve (AUC) of 0.843 and 0.811, respectively. Bioinformatics analysis revealed miR-1246, as a tumor suppressor in GC, could regulate several signaling pathways.


Circulating exosomal miR-1246 was a potential biomarker for the early diagnosis of GC.


Exosomes miR-1246 Gastric cancer Diagnosis Biomarker 


Compliance with ethical standards

Conflict of interest

The authors report no conflicts of interest in this work.

Ethical approval

This study was approved by the Research and Ethical Committee of The Second Affiliated Hospital of Nanjing Medical University.

Informed consent

Written informed consent was obtained from all participants


  1. 1.
    Van Cutsem E, Sagaert X, Topal B et al (2016) Gastric cancer. Lancet 388:2654–2664CrossRefGoogle Scholar
  2. 2.
    Veitch AM, Uedo N, Yao K et al (2015) Optimizing early upper gastrointestinal cancer detection at endoscopy. Nat Rev Gastroenterol Hepatol 12:660–667CrossRefGoogle Scholar
  3. 3.
    Carpelan-Holmstrom M, Louhimo J, Stenman UH et al (2002) CEA, CA 19–9 and CA 72–4 improve the diagnostic accuracy in gastrointestinal cancers. Anticancer Res 22:2311–2316Google Scholar
  4. 4.
    Hundahl SA, Phillips JL, Menck HR (2000) The National Cancer Data Base Report on poor survival of U.S. gastric carcinoma patients treated with gastrectomy: Fifth Edition American Joint Committee on Cancer staging, proximal disease, and the "different disease" hypothesis. Cancer 88:921–932Google Scholar
  5. 5.
    Hartgrink HH, Jansen EP, van Grieken NC et al (2009) Gastric cancer. Lancet 374:477–490CrossRefGoogle Scholar
  6. 6.
    Newton AD, Datta J, Loaiza-Bonilla A et al (2015) Neoadjuvant therapy for gastric cancer: current evidence and future directions. J Gastrointest Oncol 6:534–543Google Scholar
  7. 7.
    Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116:281–297CrossRefGoogle Scholar
  8. 8.
    Caldas C, Brenton JD (2005) Sizing up miRNAs as cancer genes. Nat Med 11:712–714CrossRefGoogle Scholar
  9. 9.
    Trams EG, Lauter CJ, Salem N Jr et al (1981) Exfoliation of membrane ecto-enzymes in the form of micro-vesicles. Biochim Biophys Acta 645:63–70CrossRefGoogle Scholar
  10. 10.
    Conigliaro A, Fontana S, Raimondo S et al (2017) Exosomes: nanocarriers of biological messages. Adv Exp Med Biol 998:23–43CrossRefGoogle Scholar
  11. 11.
    Zeng Z, Li Y, Pan Y et al (2018) Cancer-derived exosomal miR-25-3p promotes pre-metastatic niche formation by inducing vascular permeability and angiogenesis. Nat Commun 9:5395CrossRefGoogle Scholar
  12. 12.
    Naseri Z, Oskuee RK, Jaafari MR et al (2018) Exosome-mediated delivery of functionally active miRNA-142-3p inhibitor reduces tumorigenicity of breast cancer in vitro and in vivo. Int J Nanomed 13:7727–7747CrossRefGoogle Scholar
  13. 13.
    Li Z, Tao Y, Wang X et al (2018) Tumor-secreted exosomal miR-222 promotes tumor progression via regulating P27 expression and re-localization in pancreatic cancer. Cell Physiol Biochem 51:610–629CrossRefGoogle Scholar
  14. 14.
    Yoshimura A, Sawada K, Nakamura K et al (2018) Exosomal miR-99a-5p is elevated in sera of ovarian cancer patients and promotes cancer cell invasion by increasing fibronectin and vitronectin expression in neighboring peritoneal mesothelial cells. BMC Cancer 18:1065CrossRefGoogle Scholar
  15. 15.
    Gilad S, Meiri E, Yogev Y et al (2008) Serum microRNAs are promising novel biomarkers. PLoS ONE 3:e3148CrossRefGoogle Scholar
  16. 16.
    Lu J, Getz G, Miska EA et al (2005) MicroRNA expression profiles classify human cancers. Nature 435:834–838CrossRefGoogle Scholar
  17. 17.
    Zhong Y, Chen Z, Guo S et al (2017) TUG1, SPRY4-IT1, and HULC as valuable prognostic biomarkers of survival in cancer: a PRISMA-compliant meta-analysis. Medicine (Baltimore) 96:e8583CrossRefGoogle Scholar
  18. 18.
    Shin VY, Ng EK, Chan VW et al (2015) A three-miRNA signature as promising non-invasive diagnostic marker for gastric cancer. Mol Cancer 14:202CrossRefGoogle Scholar
  19. 19.
    Yuan HL, Wang T, Zhang KH (2018) MicroRNAs as potential biomarkers for diagnosis, therapy and prognosis of gastric cancer. Onco Targets Ther 11:3891–3900CrossRefGoogle Scholar
  20. 20.
    Blake JA, Christie KR, Dolan ME et al (2015) Gene ontology consortium: going forward. Nucleic Acids Res 43:D1049–D1056CrossRefGoogle Scholar
  21. 21.
    Kanehisa M, Sato Y, Kawashima M et al (2016) KEGG as a reference resource for gene and protein annotation. Nucleic Acids Res 44:D457–462CrossRefGoogle Scholar
  22. 22.
    da Huang W, Sherman BT, Lempicki RA (2009) Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources. Nat Protoc 4:44–57CrossRefGoogle Scholar
  23. 23.
    Szklarczyk D, Franceschini A, Wyder S et al (2015) STRING v10: protein-protein interaction networks, integrated over the tree of life. Nucleic Acids Res 43:D447–452CrossRefGoogle Scholar
  24. 24.
    Li XJ, Ren ZJ, Tang JH et al (2017) Exosomal MicroRNA MiR-1246 promotes cell proliferation, invasion and drug resistance by targeting CCNG2 in breast cancer. Cell Physiol Biochem 44:1741–1748CrossRefGoogle Scholar
  25. 25.
    Santangelo L, Giurato G, Cicchini C et al (2016) The RNA-binding protein SYNCRIP is a component of the hepatocyte exosomal machinery controlling MicroRNA sorting. Cell Rep 17:799–808CrossRefGoogle Scholar
  26. 26.
    Villarroya-Beltri C, Gutierrez-Vazquez C, Sanchez-Cabo F et al (2013) Sumoylated hnRNPA2B1 controls the sorting of miRNAs into exosomes through binding to specific motifs. Nat Commun 4:2980CrossRefGoogle Scholar
  27. 27.
    Liu X, Xu T, Hu X et al (2018) Elevated circulating miR-182 acts as a diagnostic biomarker for early colorectal cancer. Cancer Manag Res 10:857–865CrossRefGoogle Scholar
  28. 28.
    Zhang H, Wang J, Wang Z et al (2018) Serum miR-100 is a potential biomarker for detection and outcome prediction of glioblastoma patients. Cancer Biomark 24:43–49CrossRefGoogle Scholar
  29. 29.
    Orangi E, Motovali-Bashi M (2018) Evaluation of miRNA-9 and miRNA-34a as potential biomarkers for diagnosis of breast cancer in Iranian women. Gene 687:272–279CrossRefGoogle Scholar
  30. 30.
    Takeshita N, Hoshino I, Mori M et al (2013) Serum microRNA expression profile: miR-1246 as a novel diagnostic and prognostic biomarker for oesophageal squamous cell carcinoma. Br J Cancer 108:644–652CrossRefGoogle Scholar
  31. 31.
    Todeschini P, Salviato E, Paracchini L et al (2017) Circulating miRNA landscape identifies miR-1246 as promising diagnostic biomarker in high-grade serous ovarian carcinoma: a validation across two independent cohorts. Cancer Lett 388:320–327CrossRefGoogle Scholar
  32. 32.
    Bhagirath D, Yang TL, Bucay N et al (2018) microRNA-1246 is an exosomal biomarker for aggressive prostate cancer. Cancer Res 78:1833–1844CrossRefGoogle Scholar
  33. 33.
    Moshiri F, Salvi A, Gramantieri L et al (2018) Circulating miR-106b-3p, miR-101-3p and miR-1246 as diagnostic biomarkers of hepatocellular carcinoma. Oncotarget 9:15350–15364CrossRefGoogle Scholar
  34. 34.
    Wei C, Li Y, Huang K et al (2018) Exosomal miR-1246 in body fluids is a potential biomarker for gastrointestinal cancer. Biomark Med 12:1185–1196CrossRefGoogle Scholar
  35. 35.
    Madhavan B, Yue S, Galli U et al (2015) Combined evaluation of a panel of protein and miRNA serum-exosome biomarkers for pancreatic cancer diagnosis increases sensitivity and specificity. Int J Cancer 136:2616–2627CrossRefGoogle Scholar
  36. 36.
    Ogata-Kawata H, Izumiya M, Kurioka D et al (2014) Circulating exosomal microRNAs as biomarkers of colon cancer. PLoS ONE 9:e92921CrossRefGoogle Scholar
  37. 37.
    Machida T, Tomofuji T, Maruyama T et al (2016) miR1246 and miR4644 in salivary exosome as potential biomarkers for pancreatobiliary tract cancer. Oncol Rep 36:2375–2381CrossRefGoogle Scholar
  38. 38.
    Mo LJ, Song M, Huang QH et al (2018) Exosome-packaged miR-1246 contributes to bystander DNA damage by targeting LIG4. Br J Cancer 119:492–502CrossRefGoogle Scholar
  39. 39.
    Sakha S, Muramatsu T, Ueda K et al (2016) Exosomal microRNA miR-1246 induces cell motility and invasion through the regulation of DENND2D in oral squamous cell carcinoma. Sci Rep 6:38750CrossRefGoogle Scholar
  40. 40.
    Yuan D, Xu J, Wang J et al (2016) Extracellular miR-1246 promotes lung cancer cell proliferation and enhances radioresistance by directly targeting DR5. Oncotarget 7:32707–32722Google Scholar
  41. 41.
    Xu YF, Hannafon BN, Khatri U et al (2019) The origin of exosomal miR-1246 in human cancer cells. RNA Biol 16:770–784CrossRefGoogle Scholar
  42. 42.
    Kahlert UD, Mooney SM, Natsumeda M et al (2017) Targeting cancer stem-like cells in glioblastoma and colorectal cancer through metabolic pathways. Int J Cancer 140:10–22CrossRefGoogle Scholar
  43. 43.
    Lin SS, Peng CY, Liao YW et al (2018) miR-1246 targets CCNG2 to enhance cancer stemness and chemoresistance in oral carcinomas. Cancers (Basel) 10:272CrossRefGoogle Scholar
  44. 44.
    Zhang WC, Chin TM, Yang H et al (2016) Tumour-initiating cell-specific miR-1246 and miR-1290 expression converge to promote non-small cell lung cancer progression. Nat Commun 7:11702CrossRefGoogle Scholar
  45. 45.
    Tian Y, Jia X, Wang S et al (2014) SOX2 oncogenes amplified and operate to activate AKT signaling in gastric cancer and predict immunotherapy responsiveness. J Cancer Res Clin Oncol 140:1117–1124CrossRefGoogle Scholar
  46. 46.
    Polom K, Das K, Marrelli D et al (2019) KRAS mutation in gastric cancer and prognostication associated with microsatellite instability status. Pathol Oncol Res 25:333–340CrossRefGoogle Scholar
  47. 47.
    Jiang T, Xu X, Qiao M et al (2018) Comprehensive evaluation of NT5E/CD73 expression and its prognostic significance in distinct types of cancers. BMC Cancer 18:267CrossRefGoogle Scholar

Copyright information

© Japan Society of Clinical Oncology 2019

Authors and Affiliations

  • Yuntao  Shi
    • 1
    • 2
  • Zhonghong  Wang
    • 3
  • Xiaojuan  Zhu
    • 1
  • Ling  Chen
    • 1
  • Yilan  Ma
    • 1
  • Jiayan  Wang
    • 1
  • Xiaozhong  Yang
    • 2
    Email author
  • Zheng  Liu
    • 1
    Email author
  1. 1.Medical Center for Digestive DiseasesThe Second Affiliated Hospital of Nanjing Medical UniversityNanjingChina
  2. 2.Department of GastroenterologyThe Affiliated Huaian No. 1 People’s Hospital of Nanjing Medical UniversityHuaianChina
  3. 3.Department of GastroenterologyHuaian People’s Hospital of Hongze DistrictHuaianChina

Personalised recommendations