Plasma DCLK1 is a marker of hepatocellular carcinoma (HCC): Targeting DCLK1 prevents HCC tumor xenograft growth via a microRNA-dependent mechanism

Oncotarget. 2015 Nov 10;6(35):37200-15. doi: 10.18632/oncotarget.5808.

Abstract

Tumor stem cell marker Doublecortin-like kinase1 (DCLK1) is upregulated in several solid tumors. The role of DCLK1 in hepatocellular carcinoma (HCC) is unclear. We immunostained tissues from human livers with HCC, cirrhosis controls (CC), and non-cirrhosis controls (NCC) for DCLK1. Western blot and ELISA analyses for DCLK1 were performed with stored plasma samples. We observed increased immunoreactive DCLK1 in epithelia and stroma in HCC and CCs compared with NCCs, and observed a marked increase in plasma DCLK1 from patients with HCC compared with CC and NCC. Analysis of the Cancer Genome Atlas' HCC dataset revealed that DCLK1 is overexpressed in HCC tumors relative to adjacent normal tissues. High DCLK1-expressing cells had more epithelial-mesenchymal transition (EMT). Various tumor suppressor miRNAs were also downregulated in HCC tumors. We evaluated the effects of DCLK1 knockdown on Huh7.5-derived tumor xenograft growth. This was associated with growth arrest and a marked downregulation of cMYC, and EMT transcription factors ZEB1, ZEB2, SNAIL, and SLUG via let-7a and miR-200 miRNA-dependent mechanisms. Furthermore, upregulation of miR-143/145, a corresponding decrease in pluripotency factors OCT4, NANOG, KLF4, and LIN28, and a reduction of let-7a, miR-143/145, and miR-200-specific luciferase activity was observed. These findings suggest that the detection of elevated plasma DCLK1 may provide a cost-effective, less invasive tool for confirmation of clinical signs of cirrhosis, and a potential companion diagnostic marker for patients with cirrhosis and HCC. Our results support evaluating DCLK1 as a biomarker for detection and as a therapeutic target for eradicating HCC.

Keywords: HCC; biomarker; circulating DCLK1; cirrhosis; miRNA.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Biomarkers, Tumor / blood*
  • Biomarkers, Tumor / genetics
  • Carcinoma, Hepatocellular / blood
  • Carcinoma, Hepatocellular / enzymology
  • Carcinoma, Hepatocellular / genetics
  • Carcinoma, Hepatocellular / pathology
  • Carcinoma, Hepatocellular / therapy*
  • Cell Line, Tumor
  • Cell Proliferation
  • Databases, Genetic
  • Gene Expression Regulation, Neoplastic
  • Gene Knockdown Techniques*
  • Genetic Therapy / methods*
  • Humans
  • Intracellular Signaling Peptides and Proteins / blood*
  • Intracellular Signaling Peptides and Proteins / genetics
  • Liver Cirrhosis / blood
  • Liver Cirrhosis / enzymology
  • Liver Neoplasms / blood
  • Liver Neoplasms / enzymology
  • Liver Neoplasms / genetics
  • Liver Neoplasms / pathology
  • Liver Neoplasms / therapy*
  • Mice, Nude
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Neoplastic Stem Cells / enzymology
  • Neoplastic Stem Cells / pathology
  • Phenotype
  • Protein-Serine-Threonine Kinases / blood*
  • Protein-Serine-Threonine Kinases / genetics
  • RNA Interference
  • RNAi Therapeutics*
  • Retrospective Studies
  • Signal Transduction
  • Time Factors
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Transfection
  • Tumor Burden
  • Up-Regulation
  • Xenograft Model Antitumor Assays

Substances

  • Biomarkers, Tumor
  • Intracellular Signaling Peptides and Proteins
  • MicroRNAs
  • Transcription Factors
  • DCLK1 protein, human
  • Protein-Serine-Threonine Kinases