A human cancer xenograft model utilizing normal pancreatic duct epithelial cells conditionally transformed with defined oncogenes

Yuki Inagawa, Kenji Yamada, Takashi Yugawa, Shin Ichi Ohno, Nobuyoshi Hiraoka, Minoru Esaki, Tatsuhiro Shibata, Kazunori Aoki, Hideyuki Saya, Tohru Kiyono

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8 Citations (Scopus)

Abstract

Pancreatic ductal adenocarcinomas (PDACs) are considered to arise through neoplastic transformation of human pancreatic duct epithelial cells (HPDECs). In order to evaluate the biological significance of genetic and epigenetic alterations in PDACs, we isolated primary HPDECs and established an in vitro carcinogenesis model. Firstly, lentivirus-mediated transduction of KRASG12V, MYC and human papillomavirus 16 (HPV16) E6/E7 under the control of a tetracyclin-inducible promoter efficiently immortalized and transformed primary HPDECs, which gave rise to adenocarcinomas subcutaneously in an immune-deficient mouse xenograft model, depending on expression of the four genes. The tumors regressed promptly upon shutting-off the oncogenes, and the remaining tissues showed histological features corresponding to normal ductal structures with simple columnar epithelium. Re-expression of the oncogenes resulted in development of multiple PDACs through pancreatic intraepithelial neoplasia-like structures. We also succeeded in efficient immortalization of primary HPDECs with transduction of mutant CDK4, cyclin D1 and TERT. The cells maintained a normal diploid status and formed duct-like structures in a three dimensional culture. In combination with p53 silencing, KRASG12V alone was sufficient to fully transform the immortalized HPDECs, and MYC markedly accelerated the development of tumors. Our PDAC model supports critical roles of KRAS mutations, inactivation of the p53 and p16-pRB pathways, active telomerase and MYC expression in pancreatic carcinogenesis and thus recapitulates many features of human PDAC development. The present system with reversible control of oncogene expression enabled de novo development of PDAC from quasi-normal human tissues pre-formed subcutaneously in mice and might be applicable to carcinogenesis models in many organ sites.

Original languageEnglish
JournalCarcinogenesis
Volume35
Issue number8
DOIs
Publication statusPublished - 08-2014
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Cancer Research

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    Inagawa, Y., Yamada, K., Yugawa, T., Ohno, S. I., Hiraoka, N., Esaki, M., Shibata, T., Aoki, K., Saya, H., & Kiyono, T. (2014). A human cancer xenograft model utilizing normal pancreatic duct epithelial cells conditionally transformed with defined oncogenes. Carcinogenesis, 35(8). https://doi.org/10.1093/carcin/bgu112