One mouse model of intrahepatic cholangiocarcinoma by hydrodynamic injection of sleeping beauty vectors

Objective To establish one mouse model of intrahepatic cholangiocarcinoma (ICC) for liver cancer reseach. Methods The transposon vectors of sleeping beauty were constructed harboring the activated protein kinase B (myr-Akt) or the activated Notch1 intracellular domain (NICD1), respectively. These vectors were hydrodynamic injected through tail veil. Mice injected with saline was used as the control. Mice were sacrificed around 4 weeks. Tumors were featured by macro-and micro-histological observation. Immunohistological staining and fluorescence staing were used for cytokeratin (CK)19 and Akt cellular expression detected by the sequence Tag of hemagglutinin surface antigen determinant of influenza virus (HA)-tag. Results Compared with the control, mice in the experimental group progressed to tumor with the rate of 100% (14/14). The features of macro-and micro-histological observation indicate the formation of intraheptic cholangiocarcinoma. High percentage (64.3%) of CK19-positive staining in the malignant cells of liver confirmed the tumorigenesis of ICC. Similarly, high rate of HA-positive(42.9%) expression was observed in malignant cells. Colocalization of CK19 and HA staining by immunofluorescence staining also verify high expression. Conclusion We provide one method to generate ICC murine model with the advantages of easy-manupilation and time-saving and recapitulation. Key words: Intrahepatic cholangiocarcinoma; Hydrodynamic Injection; Sleeping beauty transposons; Mouse, model