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We have established transgenic rat lines carrying a human HrasG12V (Hras250) [4] or a human KrasG12V (Kras327) oncogene in which the BMS-354825 of the transgene is regulated by the Cre/loxP system (termed ras Tg rats). Targeted activation of the transgene is accomplished by injection of a Cre recombinase-carrying adenovirus (AxCANCre) into the pancreatic ducts through the common bile duct. Neoplastic lesions in the ras Tg rats exhibit morphological similarities to those observed in human pancreas lesions. Early ductal lesions exhibit close similarity to intraepithelial neoplasias (PanIN category).
Materials and methods
Tumor induction and pathological examination. AxCANCre was amplified in HEK293 cells and then purified using Vivapure Adenopack (Vivascience, Hannover, Germany). The titer of the adenovirus was determined using a rapid titer kit (Clontech, Mountain View, CA). Pancreas tumors were induced as described previously [4]. Ultrasound images were acquired using a microimaging system (EUB-8500, Hitachi Medical Corp., Tokyo, Japan). The pancreas was frozen in liquid nitrogen for RNA assays, or fixed in phosphate-buffered 10% formalin and processed for histological observation. Primary antibody anti-rat-C-ERC/Mesothelin (306) (IBL, Gunma, Japan) was used, and staining was performed using Vectastain ABC kits (Vector Laboratories, Burlingame, CA).