- 作者：Aamer M. Qazi ; Oksana Gruzdyn ; Assaad Semaan ; Shelly Seward ; Sreedhar Chamala ; Vasu Dhulipala ; Seema Sethi ; Rouba Ali-Fehmi ; Philip A. Philip ; David L. Bouwman ; Donald W. Weaver ; Scott A. Gruber ; Ramesh B. Batchu
- 刊名：Surgery
- 出版年：2012
- 出版时间：October, 2012
- 年：2012
- 卷：152
- 期：4
- 页码：704-713
- 全文大小：661 K
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.