Hoffman R M
AntiCancer, Inc., San Diego, CA, USA.
Biotechniques. 2001 May;30(5):1016-22, 1024-6. doi: 10.2144/01305bi01.
We have developed mouse models of metastatic cancer with genetically fluorescent tumors that can be imaged in fresh tissue, in situ, as well as externally. To achieve this capability, we have transduced the green fluorescent protein (GFP) gene, cloned from the bioluminescent jellyfish Aequorea victoria, into a series of human and rodent cancer cell lines that were selected in vitro to stably express GFP in vivo after transplantation to metastatic rodent models. Techniques were also developed for transduction of tumors by GFP in vivo. With this fluorescent tool, we detected and visualized for the first time tumors and metastasis in fresh viable tissue or in situ in host organs down to the single-cell level. GFP tumors on the colon, prostate, breast, brain, liver, lymph nodes, lung, pancreas, bone, and other organs can also be visualized externally, transcutaneously by quantitative whole-body fluorescence optical imaging. Real-time tumor and metastatic growth and angiogenesis and inhibition by representative drugs can be imaged and quantified for rapid antitumor, antimetastatic, and antiangiogenesis drug screening. The GFP-transfected tumor cells enabled a fundamental advance in the visualization of tumor growth and metastasis in real time in vivo.
我们已经构建了具有基因荧光肿瘤的转移性癌症小鼠模型，这种肿瘤在新鲜组织、原位以及体外均可成像。为实现这一功能，我们将从生物发光水母维多利亚多管水母中克隆出的绿色荧光蛋白（GFP）基因转导至一系列人和啮齿动物癌细胞系中，这些细胞系在体外经过筛选，移植到转移性啮齿动物模型后能在体内稳定表达GFP。还开发了在体内通过GFP转导肿瘤的技术。借助这种荧光工具，我们首次在新鲜的存活组织或宿主器官原位检测并可视化肿瘤及转移灶，直至单细胞水平。结肠、前列腺、乳腺、脑、肝、淋巴结、肺、胰腺、骨及其他器官上的GFP肿瘤，也可通过定量全身荧光光学成像经皮在体外进行可视化。实时肿瘤及转移灶的生长、血管生成以及代表性药物的抑制作用均可成像并定量，以用于快速的抗肿瘤、抗转移及抗血管生成药物筛选。GFP转染的肿瘤细胞使体内实时可视化肿瘤生长和转移取得了根本性进展。
