Abstract 5240: Establishment of novel human pancreatic cancer cell lines from liver and lung metastases in NOD/SCID/γc null (NOG) mice

Abstract

Abstract BACKGROUND: Pancreatic cancer is associated with extremely high mortality rates due to rapid progression and a high incidence of metastases. Liver and lung metastases in the early postoperative period are one of the causes for the poor prognosis of patients with resected pancreatic cancer. Information on the pathobiology of metastatic pancreatic cancer and elucidation of the underlying molecular mechanisms are urgently needed in order to develop novel and effective therapeutic approaches to treatment. Injection of human pancreatic cancer cells in the spleens of nude mice is one of the major liver metastasis models. However, this model yields few metastatic tumors in the livers and none in the lungs. NOG mice have severe immunodeficiency, including defects of T, B and NK cell functions, and dysfunction of dendritic cells. Human pancreatic cancer cells are reported to easily proliferate and metastasize to other organs in NOG mice. In this study, we established and characterized human pancreatic cancer cell lines from liver and lung metastases in NOG mice. METHODS: PANC-1 cells (1×105 cells) were injected into the spleens of NOG mice. The mice were euthanized 8 weeks after injection, and the livers and lungs were removed. Metastatic tumors of the liver and lung were cut into small pieces and dispersed into single cells in a medium containing antibiotics. Biological and morphological characteristics, and gene expression patterns of mesenchymal markers of the liver and lung metastatic cells, were compared with parental cells. RESULTS: These metastatic cells were confirmed as being of human-cell origin using PCR with primer pair of human mitochondrial DNA. There were no morphological changes between liver and lung metastatic cells and parental cells, but the metastatic cells proliferated more slowly than parental cells as determined by MTT assays and cell counts. By contrast, the metastatic cells exhibited greater migration than the parental cells, as determined in Boyden chamber assays and by time lapse analysis. Attachment assays revealed that adhesion of metastatic cells to the type I and IV collagen, laminin and fibronectin was stronger than that of parental cells. Nestin, a cancer stem cell marker, was highly expressed in the liver and lung metastatic cells. CONCLUSION: These findings suggest that human pancreatic cancer cell lines derived from liver and lung metastases in NOG mice have different characteristics from parental cells. These new cell lines may contribute to resolving the mechanisms of metastasis of pancreatic cancer and the roles of cancer stem cells in metastasis. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5240. doi:10.1158/1538-7445.AM2011-5240