Department of Pediatric Surgery, Children's University Hospital Tuebingen, Tuebingen, Germany.
PLoS One. 2011;6(8):e23419. doi: 10.1371/journal.pone.0023419. Epub 2011 Aug 10.
Investigation of hepatoblastoma in experimental conditions contributes relevantly to a detailed understanding of tumor biology and the investigation of new treatment approaches. Most systematical analyses currently use subcutaneous xenografts. We established a reproducible intrahepatic model with the hepatoblastoma-cell lines HuH6 and HepT1. The cells were stably transfected with a plasmid vector encoding for Gaussia luciferase. HuH6 and HepT1 were injected intrasplenically in NOD/LtSz-scid IL2Rγnull mice. Mice were splenectomized in order to avoid intrasplenical tumor growth. Multifocal intrahepatic tumor growth was observed in 85% (11/13) of HuH6 tumors and 55% (5/9) of HepT1 tumors. Serum Alpha-fetoprotein and Gaussia luciferase increased 5 weeks after tumor-cell inoculation. Tumors were detected by MRI at this time point. Immunhistochemical analysis such as vascularity (CD31), proliferation index (Ki-67), cytokeratin 7 and distribution of β-catenin in intrahepatic tumors were different to subcutaneous tumors. We established a reproducible xenograft model for intrahepatic hepatoblastoma growth with a high tumor incidence. Monitoring of tumor cell viability was optimized by measuring GLuc. This model enables further experimental investigations of HB in a more physiological milieu as emphasized by the β-catenin distribution.
在实验条件下研究肝母细胞瘤对深入了解肿瘤生物学和研究新的治疗方法有重要贡献。目前大多数系统分析都使用皮下异种移植。我们用肝母细胞瘤细胞系 HuH6 和 HepT1 建立了一种可重复的肝内模型。这些细胞被稳定地转染了一个编码 Gaussia 荧光素酶的质粒载体。HuH6 和 HepT1 被注射到 NOD/LtSz-scid IL2Rγnull 小鼠的脾内。为了避免脾内肿瘤生长,对小鼠进行了脾切除术。在 HuH6 肿瘤的 85%(11/13)和 HepT1 肿瘤的 55%(5/9)中观察到多灶性肝内肿瘤生长。在肿瘤细胞接种后 5 周,血清甲胎蛋白和 Gaussia 荧光素酶增加。此时可以通过 MRI 检测到肿瘤。肝内肿瘤的免疫组织化学分析,如血管生成(CD31)、增殖指数(Ki-67)、细胞角蛋白 7 和 β-连环蛋白的分布,与皮下肿瘤不同。我们建立了一种可重复的肝内肝母细胞瘤生长的异种移植模型,肿瘤发生率很高。通过测量 GLuc 优化了肿瘤细胞活力的监测。该模型使我们能够在更生理的环境中进一步研究 HB,β-连环蛋白的分布强调了这一点。