AntiCancer, Inc., 7917 Ostrow Street, San Diego, California, 92111; Department of Dermatology, Kitasato University School of Medicine, Kanagawa, Japan; Department of Surgery, University of California, San Diego, 200 West Arbor Drive, San Diego, California, 92103-8220.
J Cell Biochem. 2014 Mar;115(3):457-63. doi: 10.1002/jcb.24677.
Lymph nodes are often the first target of metastatic cancer which can then remetastasize to distant organs. The progression of lymph node metastasis is dependent on sufficient blood supply provided by angiogenesis. In the present study, we have developed a color-coded imaging model to visualize angiogenesis of lymph nodes metastasis using green fluorescent protein (GFP) and red fluorescent protein (RFP). Transgenic mice carrying GFP under the control of the nestin promoter (ND-GFP mice) were used as hosts. Nascent blood vessels express GFP in these mice. B16F10-RFP melanoma cells were injected into the efferent lymph vessel of the inguinal lymph node of the ND-GFP nude mice, whereby the melanoma cells trafficked to the axillary lymph node. Three days after melanoma implantation, ND-GFP-expressing nascent blood vessels were imaged in the axillary lymph nodes. Seven days after implantation, ND-GFP-expressing nascent blood vessels formed a network in the lymph nodes. ND-GFP-positive blood vessels surrounded the tumor mass by 14 days after implantation. However, by 28 days after implantation, ND-GFP expression was diminished as the blood vessels matured. Treatment with doxorubicin significantly decreased the mean nascent blood vessel length per tumor volume. These results show that the dual-color ND-GFP blood vessels/RFP-tumor model is a powerful tool to visualize and quantitate angiogenesis of metastatic lymph nodes as well as for evaluation of its inhibition.
淋巴结通常是转移性癌症的第一个靶标，然后可以转移到远处的器官。淋巴结转移的进展依赖于血管生成提供的足够的血液供应。在本研究中，我们开发了一种彩色成像模型，使用绿色荧光蛋白（GFP）和红色荧光蛋白（RFP）可视化淋巴结转移的血管生成。携带巢蛋白启动子控制下 GFP 的转基因小鼠（ND-GFP 小鼠）被用作宿主。在这些小鼠中，新生血管表达 GFP。B16F10-RFP 黑色素瘤细胞被注入腹股沟淋巴结的输出淋巴管中，从而使黑色素瘤细胞转移到腋窝淋巴结。在黑色素瘤植入后 3 天，在腋窝淋巴结中成像表达 ND-GFP 的新生血管。在植入后 7 天，ND-GFP 表达的新生血管在淋巴结中形成网络。在植入后 14 天，ND-GFP 阳性血管环绕肿瘤团块。然而，在植入后 28 天，随着血管成熟，ND-GFP 表达减少。多柔比星治疗显著降低了每肿瘤体积的平均新生血管长度。这些结果表明，双色 ND-GFP 血管/RFP-肿瘤模型是一种强大的工具，可用于可视化和定量转移性淋巴结的血管生成，并可用于评估其抑制作用。
