Abstract 1486: Role of Slug in chemoresistance, invasion and metastasis of CD133-expressing pancreatic cancer.

Abstract

Abstract Background: Pancreatic cancer is a devastating disease despite current development of therapy. Accumulating evidence suggests that cancer stem cells (CSCs) are responsible for recurrence and chemoresistance. CD133, a marker of CSCs in various solid tumors including pancreatic cancer, has been studied for recent decade. However, the role of CD133 is still obscure. Epithelial-mesenchymal transition (EMT), which is the essential process in the wound healing, development and cancer progression, is one of the CSC characteristics. However, the role of EMT in the pancreatic cancer progression is not sufficiently elucidated. Slug is a member of the Snail family of zinc finger transcription factors and plays a role in the EMT. Slug is demonstrated to be overexpressed in numerous cancers, and involved in migration and tumorigenesis. Recent study demonstrated that EMT was directly suppressed by downregulation of the transcription of Slug (Chakrabarti et al, Nat Cell Biol, 2012). In this study, we investigated the relationship between Slug and biological properties in pancreatic cancer cells. Materials and Methods: We established a highly migratory cell population with endogenous CD133high derived from Capan-1 cells by migration assay system. Over 90% of CD133high cells expressed CD133 by flow cytometer. Consequently, we established CD133knock-down and Slugknock-down by lentiviral transduction method. We used gemcitabine (GEM) for chemosensitivity assay. To examine the role of Slug, we used migration and invasion assay, wound healing assay, DNA microarray, real-time RT-PCR and Western blot. Results: 1) DNA microarray and real-time RT-PCR revealed higher expression of EMT-related genes, such as Slug and N-cadherin, in CD133high cells compared to CD133knock-down cells, suggesting that CD133knock-down related to downregulation of Slug. 2) Slugknock-down cells showed more sensitive to GEM by MTT assay compared to parental cells (CD133high cells), but no change of CD133 expression. 3) Slugknock-down cells showed less migratory and invasive abilities than CD133high cells. 4) CD133high cells showed more in vivo metastasis compared to CD133knock-down cells using immunodeficient mice. In vivo experiment using Slugknock-down cells is now ongoing. Conclusion: Our results suggest that Slug plays an important role in chemoresistance, invasion and metastasis. Citation Format: Koichirou Tsukasa, Qiang Ding, Makoto Yoshimitsu, Yumi Miyazaki, Toru Obara, Shyuichiro Matsubara, Koki Maeda, Sonshin Takao. Role of Slug in chemoresistance, invasion and metastasis of CD133-expressing pancreatic cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1486. doi:10.1158/1538-7445.AM2013-1486