Abstract 3924: Establishment and characterization of two cell lines of anaplastic pancreatic cancer
Authors: Miura Kotaro , Kimura Kenjiro , Amano Ryosuke , Yamazoe Sadaaki , Hirata Keiichiro , Shibutani Masatsune , Sakurai Katsunobu , Nagahara Hisashi , Toyokawa Takahiro , Kubo Naoshi , Tanaka Hiroaki , Muguruma Kazuya , Otani Hiroshi , Yashiro Masakazu , Maeda Kiyoshi , Ohira Masaichi , Hirakawa Kosei 
Published: May 2015

Abstract
Background: Anaplastic pancreatic cancer is a rare malignancy with a poor prognosis. As a whole, pancreatic cancer is a devastating prognosis but its therapeutic strategy has not still well developed. Thus, new models are needed to research its biology.
Purpose: We have established and characterized 2 anaplastic cancer cell lines (OCUP-A1 and OCUP-A2) for revealing tumor worsening factors and the mechanism of dedifferentiation.
Methods: We researched the present studies by comparing with established 2 cell lines and well-known pancreatic cancer cell lines. In vitro proliferation, migration and invasion assays was examined in each cell lines under normoxia and hypoxia by CellPlayer™ 96-Well Kinetic Cell Proliferation, Migration and Invasion Assays. Furthermore,as assessment of chemosensitivity, IC50 values against 5Fu and gemcitabine ware measured by MTT assay. As In vivo assays, the growth of the xenografts in each cell lines was also measured for a month. The immunohistochemistry (IHC) of E-cadherin and vimentin for primary tumors and the xenografts was performed to confirm the induction of epitherial-mesenchymal transition (EMT).
Result: Both OCUP-A1 and OCUP-A2 were pleomorphic cells derived from ascites of anaplastic pancreatic cancer patients. The doubling time of OCUP-A1 and OCUP-A2 was 30.9 h and 20.4 h, respectively. In migration and invasion assays, relative wound density in new 2 cell lines ranged from 47.1 % to 98.1 % at 48 hours. The values were not so different from that of other cell lines. Interestingly, although a large part of OCUP-A1 is spindle shape in normoxia, the subpopulation with polygonal structure has increased in hypoxia. Furthermore, OCUP-A1 rapidly proliferated in hypoxia. The IC50 values of gemcitabine for OCUP-A1 and OCUP-A2 were 8.77 nM and 4.90 nM, respectively. And the values of 5-Fu for OCUP-A1 and OCUP-A2 were 48.5 μM and 14.8μM, respectively. Chemosensitivity of 2 established cell lines was not superior to other cell lines. The xenografts of OCUP-A1 and OCUP-A2 more rapidly increased than other cell lines (MIAPaCa2 and Panc-1). In IHC, the E-cadherin expression was reduced and the vimentin expression was enhanced in both specimens.
Conclusion: We established 2 anaplastic pancreatic cancer cell lines. It was suspected that these cell lines would have aggressive characters affected by TME and EMT.
Citation Format: Kotaro Miura, Kenjiro Kimura, Ryosuke Amano, Sadaaki Yamazoe, Keiichiro Hirata, Masatsune Shibutani, Katsunobu Sakurai, Hisashi Nagahara, Takahiro Toyokawa, Naoshi Kubo, Hiroaki Tanaka, Kazuya Muguruma, Hiroshi Otani, Masakazu Yashiro, Kiyoshi Maeda, Masaichi Ohira, Kosei Hirakawa. Establishment and characterization of two cell lines of anaplastic pancreatic cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3924. doi:10.1158/1538-7445.AM2014-3924