Restoration of E-cadherin expression in pancreatic ductal adenocarcinoma treated with microRNA-101

Surgery. 2012 Oct;152(4):704-11; discussion 711-3. doi: 10.1016/j.surg.2012.07.020. Epub 2012 Sep 1.

Abstract

Purpose: To investigate the possibility of inhibiting the progression of pancreatic ductal adenocarcinoma (PDAC) by facilitating the expression of E-cadherin through the enforced expression of microRNA-101 (miR-101).

Methods: In situ hybridization was conducted with archival tissue using a double digoxigenin-labeled probe. Chromatin immunoprecipitation (ChIP) assay was conducted with EZ-Magna ChIPTM A. Gene profile analysis, Western blot, and immunoprecipitation assays were performed using standard protocols.

Results: We found that decreased miR-101 expression observed in archival patient tissues was significantly associated with poor prognosis indicated by low-intensity staining in high-grade tumors. ChIP assays using anti-enhancer of zeste homolog 2 (EZH2) antibodies indicated not only the interaction of EZH2 to the CDH1 (E-cadherin) promoter, but also that this interaction was significantly diminished in cells transfected with pre-miR-101. We observed a global downregulation of trimethylated lysine 27 of H3 histone (H3K27me3) along with upregulation of the enzymes histone deacetylase -1 and -2 with the re-expression of miR-101. Further, we observed lesser levels of transcriptional factors that inhibit the CDH1 promoter with pre-miR-101 treatment. Western blot analysis confirmed the enhanced E-cadherin expression. PANC-1 cells transduced with pre-miR-101 displayed markedly attenuated growth in SCID mice.

Conclusion: These results suggest the potential therapeutic use of miR-101-enforced expression for inhibition of PDAC.

MeSH terms

  • Adult
  • Aged
  • Animals
  • Antigens, CD
  • Apoptosis / genetics
  • Base Sequence
  • Cadherins / genetics*
  • Carcinoma, Pancreatic Ductal / genetics*
  • Carcinoma, Pancreatic Ductal / pathology
  • Carcinoma, Pancreatic Ductal / therapy*
  • Case-Control Studies
  • Cell Line, Tumor
  • Cell Movement / genetics
  • Cell Proliferation
  • Female
  • Gene Expression
  • Humans
  • In Situ Hybridization
  • Male
  • Mice
  • Mice, SCID
  • MicroRNAs / genetics
  • MicroRNAs / therapeutic use*
  • Middle Aged
  • Neoplasm Invasiveness / genetics
  • Pancreatic Neoplasms / genetics*
  • Pancreatic Neoplasms / pathology
  • Pancreatic Neoplasms / therapy*
  • Promoter Regions, Genetic
  • Transduction, Genetic
  • Tumor Stem Cell Assay
  • Xenograft Model Antitumor Assays

Substances

  • Antigens, CD
  • CDH1 protein, human
  • Cadherins
  • MIRN101 microRNA, human
  • MicroRNAs