肝癌临床标本原位移植模型的建立及其影像学评估

Establishment of Orthotopic Transplantation Model of Liver Cancer and Imaging Evaluation

目的建立基于临床肝癌手术标本的原位移植(patient-derived orthotopic xenograft PDOX)模型,研究近红外荧光(Near-infrared fluorescence,NIRF)活体成像和PET/CT在模型评估中的应用。方法将临床新鲜肝癌手术标本接种于重度免疫缺陷NPG小鼠皮下建立肝癌PDX模型,通过组织形态观察、STR分型检测和免疫组化分析对PDX模型进行评估。进一步将PDX模型肿瘤组织进行裸鼠肝原位移植建立PDOX模型,注射近红外荧光染料IR-783,通过小动物活体成像检测肿瘤的发生;尾静脉注射18F-FDG,通过小动物PET/CT观察确认肝原位肿瘤的生长。结果 STR分型结果表明PDX肿瘤的人源性特征,组织形态观察和免疫组化检测表明PDX肿瘤保持了原发肿瘤病理学特征;近红外荧光活体成像检测到肝脏肿瘤的发生;PET/CT可清晰观察到小鼠肝脏部位18F-FDG分子探针富集。结论成功建立了肝癌PDOX模型,通过小动物活体成像和PET/CT影像技术可对该模型进行评估,为肝癌的治疗和发病机制研究提供了良好的动物模型。

Object A patient-derived orthotopic xenograft PDOX model was established to study the application of near-infrared fluorescence（NIRF） in vivo imaging and PET/CT in model evaluation. Method The PDX model of hepatocellular carcinoma was established by subcutaneously injecting clinical fresh hepatocellular carcinoma（HCC） specimens into severely immunodeficient NPG mice. The PDX model was evaluated by histomorphometric observation,STR genotyping,and immunohistochemistry. PDX models were implanted in nude mice to establish PDOX models by orthotopic liver transplantation. Near-infrared fluorescent dye IR-783 was injected to detect the occurrence of tumors by in vivo imaging of small animals. 18 F-FDG was injected through the tail vein and observed by small animals PET/CT to confirm liver orthotopic tumor growth. Result The STR genotyping result showed that the human characteristics, histomorphological observation and immunohistochemical detection of PDX tumors indicated that PDX tumors retained the pathological features of primary tumors; detection of liver tumors was detected by near-infrared fluorescence in vivo imaging; PET/CT was available. The enrichment of 18 F-FDG molecular probes in the mouse liver was clearly observed. Conclusion The PDOX model of hepatocellular carcinoma was established successfully. The model can be evaluated by living animal imaging and PET/CT imaging technology. It provides a good animal model for the treatment and pathogenesis of hepatocellular carcinoma.