Abstract 294: CD133/ERK axis mediates invasion and metastasis through epithelial to mesenchymal transition in pancreatic cancer.

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Abstract Background: Over 90% of pancreatic cancer-associated mortality is due to metastasis, which involves several serial steps from disseminating of primary cancer cells to colonizing in the distant organs. CD133, a marker of cancer stem cells (CSCs) in various solid tumors including pancreatic cancer, has been studied for recent decade. However, the role of CD133 is still obscure. It has been supposed that the epithelial-mesenchymal transition (EMT) is one of CSC properties in the process of metastasis. The present study is to investigate the regulatory function of CD133 in migration and invasion of pancreatic cancer (PC). Methods: We previously reported the establishment of endogenous CD133high PC cells derived from a human PC cell line, Capan-1, at AACR 2011 Meeting. CD133high fraction was sorted by FACS, and CD133 expression was detected by real-time RT-PCR and Western blot. shRNA-CD133-GFP or shRNA-Slug-GFP was transfected into CD133high PC cells to establish CD133knock-down or Slug knock-down PC cells by lentivirus vector to knock down CD133 or Slug, resulting in 80-85% knock-down of endogenous CD133 or Slug. To examine EMT, we used migration and invasion assay, wound healing assay, DNA microarray, real-time RT-PCR and Western blot. Results: FACS analysis showed that CD133 was enriched by more than 90% in CD133high PC cells. CD133high PC cells showed migratory and invasive abilities two- to three-fold compared with parental cells or CD133knock-down PC cells. Expressions of Slug, N-cadherin and fibronectin were upregulated in the CD133high PC cells, which are characteristic components of EMT by DNA microarray and real-time RT-PCR, but these were downregulated in CD133knock-down PC cells. In addition, N-cadherin could be downregulated in Slug knock-down PC cells but no change of CD133 expression. We found that not only N-cadherin also Slug and CD133 were down regulated significantly by ERK inhibitor (U0126) administration. Further we treated the CD133high PC cells with N-cadherin neutral antibody, the wound healing speed was significantly slower than that without antibody neutralization. Conclusion: Taken together, N-cadherin plays an important role in migration and invasion. CD133/ERK axis and Slug interacted as a modulation loop on N-cadherin expression. Further study should shed new light to understand the orchestrated network associated with CD133 of pancreatic cancer metastasis. These insights on CD133 regulation could be a promising novel targeted therapy for pancreatic cancer. Citation Format: Qiang Ding, Makoto Yoshimitsu, Koichiro Tsukasa, Yumi Miyazaki, Shyuichiro Matsubara, Toru Obara, Sonshin Takao. CD133/ERK axis mediates invasion and metastasis through epithelial to mesenchymal transition in 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 294. doi:10.1158/1538-7445.AM2013-294