Circulating microRNAs, miR-939, miR-595, miR-519d and miR-494, Identify Cirrhotic Patients with HCC

PLoS One. 2015 Oct 28;10(10):e0141448. doi: 10.1371/journal.pone.0141448. eCollection 2015.

Abstract

The performance of circulating biomarkers for the diagnosis of hepatocellular carcinoma (HCC) is sub-optimal. In this study we tested circulating microRNAs as biomarkers for HCC in cirrhotic patients by performing a two stage study: a discovery phase conducted by microarray and a validation phase performed by qRT-PCR in an independent series of 118 patients. Beside miRNAs emerged from the discovery phase, miR-21, miR-221, miR-519d were also tested in the validation setting on the basis of literary and tissue findings. Deregulated microRNAs were assayed in HCC-derived cells in the intracellular compartment, cell culture supernatant and exosomal fraction. Serum and tissue microRNA levels were compared in 14 patients surgically treated for HCC. From the discovery study, it emerged that seven circulating microRNAs were differentially expressed in cirrhotic patients with and without HCC. In the validation set, miR-939, miR-595 and miR-519d were shown to differentiate cirrhotic patients with and without HCC. MiR-939 and miR-595 are independent factors for HCC. ROC curves of miR-939, miR-595 and miR-519d displayed that AUC was higher than AFP. An exosomal secretion of miR-519d, miR-21, miR-221 and miR-1228 and a correlation between circulating and tissue levels of miR-519d, miR-494 and miR-21 were found in HCC patients. Therefore, we show that circulating microRNAs deserve attention as non-invasive biomarkers in the diagnostic setting of HCC and that exosomal secretion contributes to discharging a subset of microRNAs into the extracellular compartment.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Aged, 80 and over
  • Biomarkers
  • Carcinoma, Hepatocellular / etiology*
  • Carcinoma, Hepatocellular / pathology
  • Cell Line, Tumor
  • Cluster Analysis
  • Exosomes
  • Female
  • Gene Expression Profiling
  • Humans
  • Liver Cirrhosis / blood
  • Liver Cirrhosis / complications*
  • Liver Cirrhosis / genetics*
  • Liver Neoplasms / etiology*
  • Liver Neoplasms / pathology
  • Male
  • MicroRNAs / blood
  • MicroRNAs / genetics*
  • Middle Aged
  • Neoplasm Staging
  • ROC Curve
  • Reproducibility of Results

Substances

  • Biomarkers
  • MIRN494 microRNA, human
  • MIRN519 microRNA, human
  • MIRN939 microRNA, human
  • MicroRNAs

Grants and funding

This study was supported by Programma di Ricerca Regione-Università 2010-2012, Regione Emilia-Romagna, Bando "Ricerca Innovativa", to LB and LG, "Innovative approaches to the diagnosis and pharmacogenetic-based therapies of primary hepatic tumors, peripheral B and T-cell lymphomas and lymphoblastic leukaemias”; Fondazione del Monte di Bologna e Ravenna, to LB, GFS and LG; Italian Ministry of University and Research – PRIN 2010-2011 to LB; Italian Association for Cancer Research (AIRC) to MN and MF; Programma di Ricerca Regione-Università 2010-2012, Regione Emilia-Romagna, Bando "Alessandro Liberati", to FF, "Identification of innovative microRNAs-based biomarkers and anti-cancer strategies for the treatment of Hepatocellular carcinoma”.