title: Restoration of E-cadherin expression in pancreatic ductal adenocarcinoma treated with microRNA-101.
abstract: OBJECTIVE
  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.
  
  
  CONCLUSIONS
  These results suggest the potential therapeutic use of miR-101-enforced expression for inhibition of PDAC.
authors.affiliation: Laboratory of Surgical Oncology & Developmental Therapeutics, Department of Surgery, Wayne State University School of Medicine, Detroit, MI 48201, USA.
authors.forename: Aamer M
authors.initials: AM
authors.lastname: Qazi
authors.forename: Oksana
authors.initials: O
authors.lastname: Gruzdyn
authors.forename: Assaad
authors.initials: A
authors.lastname: Semaan
authors.forename: Shelly
authors.initials: S
authors.lastname: Seward
authors.forename: Sreedhar
authors.initials: S
authors.lastname: Chamala
authors.forename: Vasu
authors.initials: V
authors.lastname: Dhulipala
authors.forename: Seema
authors.initials: S
authors.lastname: Sethi
authors.forename: Rouba
authors.initials: R
authors.lastname: Ali-Fehmi
authors.forename: Philip A
authors.initials: PA
authors.lastname: Philip
authors.forename: David L
authors.initials: DL
authors.lastname: Bouwman
authors.forename: Donald W
authors.initials: DW
authors.lastname: Weaver
authors.forename: Scott A
authors.initials: SA
authors.lastname: Gruber
authors.forename: Ramesh B
authors.initials: RB
authors.lastname: Batchu
mesh_terms: D000328:Adult; D000368:Aged; D000818:Animals; D015703:Antigens, CD; D017209:Apoptosis / Q000235:genetics; D001483:Base Sequence; D015820:Cadherins / Q000235:genetics*; D021441:Carcinoma, Pancreatic Ductal / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D016022:Case-Control Studies; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation; D005260:Female; D015870:Gene Expression; D006801:Humans; D017403:In Situ Hybridization; D008297:Male; D051379:Mice; D016513:Mice, SCID; D035683:MicroRNAs / Q000235:genetics / Q000627:therapeutic use*; D008875:Middle Aged; D009361:Neoplasm Invasiveness / Q000235:genetics; D010190:Pancreatic Neoplasms / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D011401:Promoter Regions, Genetic; D014161:Transduction, Genetic; D014410:Tumor Stem Cell Assay; D023041:Xenograft Model Antitumor Assays
chemical_list: D015703:Antigens, CD; C513384:CDH1 protein, human; D015820:Cadherins; C534072:MIRN101 microRNA, human; D035683:MicroRNAs