Abstract 5200: Nestin regulates stem cell functions of pancreatic cancer cells

Abstract

Abstract Introduction: Nestin, a class VI intermediate filament, is expressed in pancreatic exocrine progenitor cells. Previously, we reported that 30% of PDAC cases expressed nestin in cancer cells, and its expression was correlated with invasion (Kawamoto, et al. Hum Pathol 2009). Knockdown of nestin using shRNA in human PDAC cell lines inhibited cell migration and invasion in vitro and liver metastasis in vivo. Restoring the expression level of nestin returned the inhibition of cell migration and invasion to the previous level (Matsuda, et al. Cancer Biol Ther 2011). In this study we analyzed the expression and roles of nestin in pancreatic cancer stem cells. Methods: Expressions of stem cell markers and nestin were analyzed using immunocytochemistry and real-time PCR. To assess cancer stem cells, a sphere formation assay was performed as follows: cells were grown in an ultra-low attachment surface plate supplemented with epithelial growth factor and fibroblast growth factor 2. Nestin-regulating molecules were clarified by DNA microarray analysis. Results: The expression level of nestin was high in sphere-forming PDAC cells, and its expression levels correlated with sphere-forming ability. In DNA microarray analysis using nestin shRNA-transfected clones and nestin-expression vector transfected-clones, at a 2-fold cut-off, only transcriptional factor 4 (TCF4) showed a compatible change coincident with nestin expression. It has been reported that TCF4 is involved in the Wnt pathway, and negatively regulates the transcription of NANOG. In real-time PCR, expression levels of TCF4 and NANOG correlated with nestin expression level. Conclusion: Nestin expression levels correlated with sphere-forming ability and expression levels of TCF4 and NANOG. These results indicate that nestin regulate stemness via TCF4 and NANOG.The expression of nestin is associated with highly metastatic cancer stem-like cells, and nestin regulates migration, invasion, and metastasis by altering the pancreatic cancer stem cell function. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5200. doi:1538-7445.AM2012-5200