基于鼻咽癌潜在靶标的活体光学成像与靶向治疗研究

发布时间：2018-04-27 19:33

本文选题：人鼻咽癌 + 生物标志物　；参考：《华中科技大学》2012年博士论文

【摘要】：鼻咽癌是具有地域特性的恶性肿瘤疾病，高发于我国广东，因此又称“广东瘤”。由于其发生部位较小且隐蔽，又与重要器官相邻，所以症状多变或不明显，导致漏诊误诊，常拖至晚期才被诊断出，而晚期的治疗效果差，5年存活率极低，严重的影响到人类健康。因此迫切需要寻找新的鼻咽癌生物标志物，进而针对该生物标志物进行相应的靶向诊疗研究。生物相容性好及靶向性高的仿生纳米载体是实现特异性地杀伤肿瘤而避免其它正常组织不受药物毒害的有效手段之一。为了更好地对鼻咽癌进行研究，需要迫切地建立起可视化鼻咽癌动物模型，能实时动态地监测鼻咽癌的发生发展和转移，进而有效地评估鼻咽癌的诊疗效果。本文旨在寻找出鼻咽癌潜在的生物标志物，建立高信噪比的可视化鼻咽癌动物模型，进而借由高密度脂蛋白仿生纳米载体对鼻咽癌进行有效的靶向诊疗。得出以下结果：（1）发现在鼻咽癌细胞和鼻咽癌患者病理组织中均高表达跨膜蛋白B类1型清道夫受体（SR-B1），故将SR-B1定义为鼻咽癌潜在的生物标志物。研究发现高表达SR-B1的鼻咽癌细胞，侧向迁移能力增强，而纵向侵袭能力受抑制。SR-B1的表达量与癌细胞的生长速度和成瘤性无关。（2）发现仿高密度脂蛋白纳米颗粒（HPPS）能够特异性地识别SR-B1高表达的鼻咽肿瘤，通过抑制肿瘤细胞的侧向迁移能力和克隆形成率，进而显著性地延缓鼻咽癌在活动物体内的生长速度。（3）尝试将携带chol-si-bmi1的HPPS用于鼻咽癌的靶向基因治疗，细胞水平实验结果表明，可以有效地抑制鼻咽癌的侧向迁移能力和克隆形成率，然而动物水平的实验未获得明显的肿瘤生长抑制效果。（4）建立了一系列荧光蛋白标记的可视化鼻咽癌实验动物模型，为鼻咽癌的活体光学成像和靶向治疗提供了良好的动物模型。其中获得了超亮远红荧光蛋白mLumin标记的鼻咽癌5-8F细胞，具有良好的皮下成瘤性和转移性。同时，还获得了一株不成瘤的远红荧光蛋白Katushka S158A标记鼻咽癌细胞株5-8F-KattushkaS158A。研究证实，，这些细胞株的成瘤性与其侧群细胞比率密切相关。（5）建立了基于疱疹病毒1胸苷激酶(TK)、远红荧光蛋白（mLumin）和萤火虫荧光素酶（Fluc）的三融合报告基因的可视化鼻咽癌实验动物模型，可同时用于荧光成像、生物发光成像和microPET成像。由于将远红荧光蛋白引入到三融合报告基因中，明显提升了活体荧光成像的检测能力，从而为肿瘤的多模式成像研究提供了良好的多标记动物模型。本文发现了SR-B1为鼻咽癌的新型潜在生物标志物，为鼻咽癌的靶向诊疗提供了有效的新型膜蛋白运输靶点。证明仿高密度脂蛋白纳米颗粒HPPS能有效的延缓鼻咽癌生长，且能有效的携带siRNA对鼻咽癌细胞进行胞浆释放进而达到基因治疗的效果。该研究建立的一系列可视化鼻咽癌动物模型，为动态监测鼻咽癌的发生发展和转移，有效的药效评估和直观的评判诊疗手段提供了便利。
[Abstract]:Nasopharyngeal carcinoma (NPC) is a malignant tumor with regional characteristics, which is often called "Guangdong tumor" in Guangdong. Therefore, it is also called "Guangdong tumor". Because of its small and hidden location and adjacent to important organs, the symptoms are changeable or inconspicuous, resulting in misdiagnosis and misdiagnosis. The late treatment is poor and the survival rate of 5 years is very low, and the survival rate is very low and serious. Therefore, it is urgent to find a new biomarker for nasopharyngeal carcinoma and to study the biomarker for the target diagnosis and treatment. The biomimetic nanoscale carrier with good biocompatibility and high targeting is one of the effective means to kill the tumor specifically and avoid the drug toxicity of other normal tissues. In order to study nasopharyngeal carcinoma better, it is necessary to establish an animal model of visual nasopharyngeal carcinoma urgently. It can monitor the occurrence, development and metastasis of nasopharyngeal carcinoma in real time and dynamically, and then effectively evaluate the diagnosis and treatment effect of nasopharyngeal carcinoma.
This paper aims to find out the potential biomarkers of nasopharyngeal carcinoma and establish a visual nasopharyngeal carcinoma model with high signal-to-noise ratio, and then the effective target diagnosis and treatment of nasopharyngeal carcinoma by high density lipoprotein biomimetic nanoscale carrier. The following results are obtained:
(1) the high expression of transmembrane protein B 1 scavenger receptor (SR-B1) was found in nasopharyngeal and nasopharyngeal carcinoma patients, so SR-B1 was defined as a potential biomarker of nasopharyngeal carcinoma. The study found that nasopharyngeal carcinoma cells with high expression of SR-B1 increased the lateral migration ability and the longitudinal invasion ability was inhibited by the expression of.SR-B1 and cancer cells. The growth rate has nothing to do with the tumorigenicity.
(2) it is found that the imitated high density lipoprotein nanoparticles (HPPS) can specifically identify the high expression of SR-B1 in nasopharyngeal tumors. By inhibiting the lateral migration ability and the clone formation rate of tumor cells, the growth rate of nasopharyngeal carcinoma in active objects can be slowed down significantly.
(3) chol-si-bmi1 HPPS was used as a target gene therapy for nasopharyngeal carcinoma. The results of cell level experiment showed that the lateral migration ability and the clone formation rate of nasopharyngeal carcinoma could be effectively suppressed. However, the experiment of animal level did not obtain the obvious inhibitory effect on tumor growth.
(4) a series of fluorescent protein labeled nasopharyngeal carcinoma experimental animal models were set up to provide a good animal model for the living optical imaging and targeting therapy of nasopharyngeal carcinoma. The 5-8F cells of nasopharyngeal carcinoma labeled with ultra bright red fluorescent protein mLumin were obtained, and a good subcutaneous tumorigenicity and metastasis were obtained. The study of nasopharyngeal carcinoma cell line 5-8F-KattushkaS158A. by Katushka S158A labeled nasopharyngeal carcinoma cell line, which was not a tumor, confirmed that the tumorigenicity of these cell lines was closely related to the ratio of side group cells.
(5) a visual nasopharyngeal carcinoma experimental animal model based on the three fusion reporter of herpes virus 1 thymidine kinase (TK), far red fluorescent protein (mLumin) and firefly luciferase (Fluc), which can be used for fluorescence imaging, bioluminescence imaging and microPET imaging, is also used for the introduction of far erythrofic protein into the three fusion reporter gene. It enhances the detection ability of live fluorescence imaging, thus providing a good multi label animal model for tumor multimode imaging research.
In this paper, SR-B1 is a new potential biomarker for nasopharyngeal carcinoma, which provides a new target for transport of membrane protein for nasopharyngeal carcinoma. It is proved that HPPS can effectively delay the growth of nasopharyngeal carcinoma and can effectively carry siRNA to the cytoplasm release of nasopharyngeal carcinoma cells to achieve gene therapy. A series of visualized animal models of nasopharyngeal carcinoma have been established to facilitate the dynamic monitoring of the occurrence, development and metastasis of nasopharyngeal carcinoma, effective drug effectiveness evaluation and intuitionistic evaluation of diagnosis and treatment.

【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R739.63

【参考文献】