CLARITY技术构建树鼩肝癌血管三维成像及相关研究

项目名称： CLARITY技术构建树鼩肝癌血管三维成像及相关研究

项目编号： No.81460426

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 邬国斌

作者单位： 广西壮族自治区肿瘤防治研究所

项目金额： 47万元

中文摘要： 新生血管形成在肝癌发生发展以及转移过程中起着关键作用，而目前肝癌血管研究局限在二维层面，无法体现肝癌血管形成过程空间结构变化及精确测量，此外肝癌血管调控机制等信息也无法直接从完整组织中获取。CLARITY技术解决了这些技术难题。CLARITY技术构建可渗透大分子并保留生物遗传信息的透明完整组织模型，免疫荧光标记后运用激光共聚焦显微镜实现研究对象三维成像，为肝癌血管形成研究提供了一个全新的技术手段。本研究以树鼩为研究对象，运用CLARITY技术构建黄曲霉毒素B1诱导的树鼩肝癌形成各个阶段肝脏透明模型，免疫荧光标记血管内皮细胞CD34和血管生成因子VEGF后，运用激光共聚焦显微镜三维成像。了解肝癌血管形成过程三维结构变化并量化及与血管生成调控因子的相互关系，为肝癌治疗靶点提供新的线索。

中文关键词： CLARITY；技术；三维图像；肝细胞癌；新生血管生成

英文摘要： Angiogenesis plays a key role in hepatocarcinogenesis and metastasis. At present, studies of angiogenesis are limited in two-dimensional level, so the changes of the vascular spatial structure are not able to be shown correctly and measured accurately. Moreover, information on the regulatory mechanism of angiogenesis is not able to be obtained directly from intact tissue either. A new technology named CLARITY solves these technical problems. CLARITY technology creates an intact tissue that is permeable to both visible-spectrum photons and exogenous macromolecules and preserves genetic information. After immunofluorescence, interesting biologic information is formed to three-dimensional imaging by confocal microscopy. In our study, some tree shrews are induced hepatocarcinogenesis by feeding aflatoxin B1. This tree shrew's livers are made transparent and permeable by CLARITY technology, and then are marked CD34 and VEGF by immunofluorescence. Finally Angiogenesis in HCC is formed to three-dimensional imaging by confocal microscopy. The three-dimensional changes of angiogenesis will be learned and measured accurately, the relation between angiogenesis and regulating factor will be investigated also. It will provide a new clue for treatment targets to cure hepatocellular carcinoma.

英文关键词： CLARITY technology;third dimension imaging;hepatocellular carcinoma vascular;angiogenesis