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

• Published in: Surgery 2012-10, Vol.152 (4), p.704-713
• Main Authors: Qazi, Aamer M., PhD, Gruzdyn, Oksana, BS, Semaan, Assaad, MD, Seward, Shelly, MD, Chamala, Sreedhar, MD, Dhulipala, Vasu, MD, Sethi, Seema, MD, Ali-Fehmi, Rouba, MD, Philip, Philip A., MD, Bouwman, David L., MD, Weaver, Donald W., MD, Gruber, Scott A., MD, PhD, MBA, Batchu, Ramesh B., PhD
• Format: Article
• Language: English
• Subjects: Adult; Aged; Animals; Apoptosis - genetics; Base Sequence; Biological and medical sciences; 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; Gastroenterology. Liver. Pancreas. Abdomen; Gene Expression; General aspects; Humans; In Situ Hybridization; Liver. Biliary tract. Portal circulation. Exocrine pancreas; Male; Medical sciences; 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; Surgery; Transduction, Genetic; Tumor Stem Cell Assay; Tumors; Xenograft Model Antitumor Assays
• ISSN: 0039-6060; 1532-7361
• DOI: 10.1016/j.surg.2012.07.020

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.