Abstract 2032: MicroRNA control of tumor cell plasticity in a new tumor metastasis model

Abstract

Abstract We have developed a new prostate cancer model to discover the molecular changes underlying metastasis. A series of cell lines were developed by isolating metastatic cells from lymph nodes of nude mice following orthotopic injection of DU145 human prostate cancer cells and expression changes analyzed using gene chips. We found that these DU145-LN cells exhibit a mesenchymal-epithelial transition (MET). Repeated rounds of in vivo metastatic selection progressively increased the expression of epithelial markers including E-cadherin, cytokeratin-18 and EpCAM, and decreased mesenchymal markers such as vimentin. Despite being more epithelial, DU145-LN cells are more migratory and show reduced proliferation in vitro. However, they show increased tumorigenesis when reinjected into nude mice. Interestingly, clinical metastases are frequently epithelial, despite strong evidence that EMT enhances metastasis. To explain this, current models of metastasis invoke EMT with subsequent MET at the secondary site. Our cells appear to demonstrate this plasticity making them an ideal model system for studying MET regulation. The MET changes in the DU145-LN cells suggested a coordinated mechanism of regulation. MicroRNA arrays revealed several candidates which correlated with the epithelial phenotype. DU145-LN cells showed increased expression of the miR-200 family (a, b, c and miR-141), and treatment with synthetic miR-200 inhibitors resulted in reduced cell-cell interactions and decreased E-cadherin levels. Our data complement recent evidence showing down-regulation of miR-200 by EMT. miR-200 acts in a feedback loop with the transcription factor ZEB1 to regulate EMT. We also observed significantly reduced expression of an additional microRNA not previously linked to cell morphology in the DU145-LN cells. Transfection of precursor miRNA to this target altered the epithelial morphology and reduced E-cadherin levels, suggesting an additional pathway of MET/EMT control. We are currently determining the mechanism of action of this new microRNA target. Our data support a role for MET in tumor metastasis, and indicate that microRNAs are an important mechanism mediating tumor cell plasticity and EMT/MET. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2032.