Whole-body and intravital optical imaging of angiogenesis in orthotopically implanted tumors
Whole-body and intravital optical imaging of angiogenesis in orthotopically implanted tumors
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DOI：
10.1073/pnas.051626698
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发表时间：
2001-02-27
影响因子：
11.1
通讯作者：
Hoffman, RM
中科院分区：
文献类型：
--
作者：
Yang, M;Baranov, E;Hoffman, RM
为了开发控制肿瘤血管生成的药物，需要一种简单、准确、经济的肿瘤诱导血管生成测定方法。我们将维多利亚绿荧光蛋白标记的人肿瘤移植到裸鼠体内，使原位植入模型适应血管生成的测量。非发光诱导毛细血管在非常明亮的肿瘤荧光中清晰可见，无论是通过活体检查还是通过全身实时亮度检查。人类癌症原位植入模型已经很好地表征，荧光阴影取代了测定血管密度的费力的组织学技术。原位植入的人胰腺肿瘤的活体图像清楚地显示原发和转移部位的血管生成毛细血管。在19天的时间内，对单个裸鼠原位生长的人前列腺肿瘤进行了血管生成的定量时间过程。通过在裸鼠足垫s.c.部位注射荧光Lewis肺癌细胞，观察肿瘤血管生成的全身光学成像。足垫相对透明，常驻血管相对较少，可对完整动物的肿瘤血管生成进行定量成像。全身成像显示，毛细血管密度在10天内呈线性增加。同样，将表达绿色荧光蛋白的人乳腺肿瘤MDA-MB-435原位移植到小鼠脂肪垫中，全身光学成像显示血管密度在20周内线性增加。这些功能强大且具有临床相关性的血管生成小鼠模型可用于生理微环境中抑制或促进肿瘤血管生成的药物的实时体内评估。
The development of drugs for the control of tumor angiogenesis requires a simple, accurate, and economical assay for tumor-induced vascularization. We have adapted the orthotopic implantation model to angiogenesis measurement by using human tumors labeled with Aequorea victoria green fluorescent protein for grafting into nude mice. The nonluminous induced capillaries are clearly visible against the very bright tumor fluorescence examined either intravitally or by whole-body luminance in real time. The orthotopic implantation model of human cancer has been well characterized, and fluorescence shadowing replaces the laborious histological techniques for determining blood vessel density. Intravital images of orthotopically implanted human pancreatic tumors clearly show angiogenic capillaries at both primary and metastatic sites. A quantitative time course of angiogenesis was determined for an orthotopically growing human prostate tumor periodically imaged intravitally in a single nude mouse over a 19-day period. Whole-body optical imaging of tumor angiogenesis was demonstrated by injecting fluorescent Lewis lung carcinoma cells into the s.c. site of the footpad of nude mice. The footpad is relatively transparent, with comparatively few resident blood vessels, allowing quantitative imaging of tumor angiogenesis in the intact animal. Capillary density increased linearly over a 10-day period as determined by whole-body imaging. Similarly, the green fluorescent protein-expressing human breast tumor MDA-MB-435 was orthotopically transplanted to the mouse fat pad, where whole-body optical imaging showed that blood vessel density increased linearly over a 20-week period. These powerful and clinically relevant angiogenesis mouse models can be used for real-time in vivo evaluation of agents inhibiting or promoting tumor angiogenesis in physiological microenvironments.