可实时监测的小鼠原发性肝癌模型建立及应用

Establishment and application of a mouse model of HCC in real-time

目的构建可实时监测的小鼠原发性肝癌(HCC)模型。方法通过高压水动力学转染技术,PB转座子系统和小鼠活体成像技术建立可实时监测的、由特定癌基因驱动的小鼠原发性肝癌模型。结果YAP5SA基因诱导表达的小鼠原发性肝癌可以成功模拟Hippo信号通路失活的肝癌亚型;通过每隔1周连续检测,整合YAP5SA-IRES-Fluc的小鼠荧光峰值信号在早期逐渐降低,于3周左右,荧光峰值信号与对照组相比,出现明显差异;PTEN-YAP5SA+模型组的荧光信号峰值与YAP5SA+对照组的荧光信号峰值相比,整体均值偏高;索拉菲尼给药1周后,本模型小鼠的荧光信号停止上升,呈现平稳的趋势。结论成功构建了一种可实时监测的小鼠原发性肝癌模型,可以进行不同肝癌亚型的分子表征,并以此为基础,为临床筛选肝癌靶向药物提供新方法。

Objective To establish a real-time monitoring mouse model for primary liver cancer(HCC)research.Methods We applied an approach for stable gene expression in mouse hepatocytes by hydrodynamic injection in combination with PiggyBac transposon system-mediated somatic gene integration.We fused an oncogene with a luciferase reporter gene for live imaging of mouse hepatocytes.Results Mouse hepatocytes integrated with a YAP1 mutant gene(YAP5SA)were used to form liver tumors with characteristics similar to the Hippo signaling pathway-inactivated HCC subclass.Live imaging of the liver showed strong a fluorescence signal from hepatocytes integrated with the oncogene and luciferase.The peak fluorescence signal decreased rapidly in the early stage after hydrodynamic injection because of hepatocyte apoptosis.The fluorescence signal was significantly increased after tumor initiation and progression.We applied this real-time monitoring model to confirm that PTEN was a HCC tumor suppressor gene by an additionally fused oncogene with shRNA expression elements.We also validated small molecular targeted drug sorafenib by oral administration.Conclusions A real-time monitoring mouse HCC model was successfully established.This method can be applied to molecular characterization of HCC subtypes and targeted drug screening.