DcR3, TFF3, and Midkine Are Novel Serum Biomarkers in Small Intestinal Neuroendocrine Tumors

Neuroendocrinology. 2017;105(2):170-181. doi: 10.1159/000452891. Epub 2016 Nov 9.


Small intestinal neuroendocrine tumors (SI-NETs) are amine- and peptide-producing neoplasms. Most patients display metastases at the time of diagnosis; they have an unpredictable individual disease course and the tumors are often therapy resistant. Chromogranin A and 5-hydroxyindoleacetic acid are the biomarkers clinically used most often today, but there is a great need for novel diagnostic and prognostic biomarkers and new therapeutic targets. Sixty-nine biomarkers were screened in serum from 23 SI-NET patients and 23 healthy controls using the multiplex proximity ligation assay (PLA). A refined method, the proximity extension assay (PEA), was used to analyze 76 additional biomarkers. Statistical testing and multivariate classification were performed. Immunohistochemistry and ELISA were performed in an extended cohort. Using PLA, 19 biomarkers showed a significant difference in serum concentrations between patients and controls, and PEA revealed a difference in the concentrations of 17 proteins. Multivariate classification analysis revealed decoy receptor 3 (DcR3), trefoil factor 3 (TFF3), and midkine to be good biomarkers for the disease, which was confirmed by ELISA analysis. All 3 biomarkers were expressed in tumor tissue. DcR3 concentrations were elevated in patients with stage IV disease. High concentrations of DcR3 and TFF3 were correlated to poor survival. DcR3, TFF3, and midkine exhibited elevated serum concentrations in SI-NET patients compared to healthy controls, and DcR3 and TFF3 were associated with poor survival. DcR3 seems to be a marker for liver metastases, while TFF3 and midkine may be new diagnostic biomarkers for SI-NETs.

Keywords: Biomarkers; Decoy receptor 3; Midkine; Proximity extension assay; Proximity ligation assay; Small intestinal neuroendocrine tumor; Trefoil factor 3.

MeSH terms

  • Biomarkers, Tumor / blood
  • Cytokines / blood*
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Follow-Up Studies
  • Humans
  • Immunohistochemistry
  • Intestinal Neoplasms / blood*
  • Intestinal Neoplasms / pathology
  • Intestine, Small / pathology
  • Liver Neoplasms / blood
  • Liver Neoplasms / secondary
  • Male
  • Middle Aged
  • Midkine
  • Multivariate Analysis
  • Neoplasm Staging
  • Neuroendocrine Tumors / blood*
  • Neuroendocrine Tumors / pathology
  • Prognosis
  • Receptors, Tumor Necrosis Factor, Member 6b / blood*
  • Survival Analysis
  • Trefoil Factor-3 / blood*


  • Biomarkers, Tumor
  • Cytokines
  • Receptors, Tumor Necrosis Factor, Member 6b
  • TFF3 protein, human
  • TNFRSF6B protein, human
  • Trefoil Factor-3
  • Midkine