Amoh Yasuyuki, Nagakura Chisa, Maitra Anirban, Moossa A R, Katsuoka Kensei, Hoffman Robert M, Bouvet Michael
AntiCancer, Inc., 7917 Ostrow Street, San Diego, California, USA.
Anticancer Res. 2006 Sep-Oct;26(5A):3237-42.
As previously shown, the stem cell marker nestin is expressed in nascent blood vessels in transgenic nestin-driven green fluorescent protein (ND-GFP) nude mice. This mouse model was recently utilized to evaluate angiogenesis in primary tumors in an orthotopic model of pancreatic cancer. In the present study, nascent angiogenesis of pancreatic cancer liver metastasis in the ND-GFP transgenic nude mice after splenic injection of low-passage xPA-1 human pancreatic cancer cells expressing red fluorescent protein (RFP) was visualized by dual-color fluorescence imaging. ND-GFP was highly expressed in proliferating endothelial cells and nascent blood vessels in the growing liver metastasis. Immunohistochemical staining showed that CD31 co-localized in ND-GFP-expressing nascent blood vessels. The density of nascent blood vessels in the tumor was readily quantitated. Gemcitabine significantly decreased the mean nascent blood vessel density in the pancreatic liver metastases. In conclusion, the dual-color model of the ND-GFP nude mouse with RFP-expressing pancreatic cancer liver metastases, enabled the simultaneous visualization and quantitation of nascent angiogenesis and its response to angiogenesis inhibitors. This model will be useful for understanding the mechanism of angiogenesis of pancreatic cancer liver metastasis and for the discovery of effective new inhibitors of this process.
如前所示，干细胞标志物巢蛋白在转基因巢蛋白驱动的绿色荧光蛋白（ND-GFP）裸鼠的新生血管中表达。最近利用该小鼠模型在胰腺癌原位模型中评估原发性肿瘤中的血管生成。在本研究中，通过双色荧光成像观察了在脾内注射表达红色荧光蛋白（RFP）的低传代xPA-1人胰腺癌细胞后，ND-GFP转基因裸鼠中胰腺癌肝转移灶的新生血管生成情况。ND-GFP在生长中的肝转移灶中的增殖内皮细胞和新生血管中高表达。免疫组织化学染色显示CD31与表达ND-GFP的新生血管共定位。肿瘤中新生血管的密度易于定量。吉西他滨显著降低了胰腺癌肝转移灶中的平均新生血管密度。总之，带有表达RFP的胰腺癌肝转移灶的ND-GFP裸鼠双色模型能够同时可视化和定量新生血管生成及其对血管生成抑制剂的反应。该模型将有助于理解胰腺癌肝转移的血管生成机制，并有助于发现针对这一过程的有效新型抑制剂。
