Abstract 2349: Establishment of a novel metastatic head and neck cancer model with gain of epithelial-mesenchymal transition and stemness properties

Abstract

Abstract Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer in the world, and includes oral squamous cell carcinoma (OSCC), which is the fourth leading cause of male cancer death in Taiwan. Metastasis is the end of neoplasm progression and the major cause of mortality of HNSCC. Prevention or elimination of metastasis may improve the morbidity and mortality caused from HNSCC. However, systemic cellular model for investigation on the metastasis of oral cancer is unavailable at present. To compromise the deviations between cell lines, we established a series of cell lines derived from an established OSCC cell line, SAS cells, which possess activity of low-tumorigenicity (SAS), highly-tumorigenicity (SASVO3 (Chen et al., J Biomed Sci 16:100, 2009)) or metastasis (SASVO3M-1 and SASVO3M-5). Metastatic SASVO3M-1 and M-5 cells were generated by i.v. injection of SASVO3 cells into the tail vein of nude mice, and the metastatic tumor in lung was collected, then, the isolated tumor cells were grown for further studies. The invasiveness activities of SASVO3M-1 and M-5 cells in lung tumor were confirmed by both in vivo and in vitro pathological examination. Metastatic cells possess increased activities of foci formation, filopodia formation, and wound healing. In the mean time, the signature of epithelial-mesenchymal transition (EMT) properties, including upregulation of transcriptional factor (Slug) and downregulation of E-cadherin, was observed. Further, metastatic cells (SASVO3M-1/-5) enhanced tumor formation in lung and shortened life span of tumor bearing mice. Next, the transcriptome of above cells was determined and then subjected to pathway analysis (Ingenuity Pathway Analysis). The group of genes specifically up-regulated in metastatic cells was centered to ERK1/2, interleukin 1α, and focal adhesion kinase. The protein level of integrin, uPAR and phosphorylated FAK was augmented in metastatic cells by immunoblot analyses. Lines of evidence support that gain of EMT properties enhances the stemness properties of cancer cells. We thus determined the stemness properties of the above three cell lines. Sphere-formation activity and the expression of stem cell marker, CD 133, were significantly induced in metastatic SASVO3M-1 and M-5 cells indicating an increase in stemness of these cells. Taken together, our data show enhanced epithelial-mesenchymal transition and stemness properties associate with the development of metastatic activity of SASVO3M-1 and M-5 cells. Further, this metastatic cell line would be an instrumental tool for further investigations on the development of diagnostic biomarkers or therapeutic targets on HNSCC. 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 2349. doi:1538-7445.AM2012-2349