Hemostasis System in Tumor Growth and Metastasis

先前的研究已经确定了止血与肿瘤生长和转移之间的关系。临床上，这些蛋白质的血浆水平升高已被认为是复发和生存率降低的预后标志物。虽然存在这些相关性，但该系统导致疾病病理学的具体体内机制在很大程度上仍然未知。几项临床研究已经确定了止血系统的组分在调节许多癌症的病理学中的作用。虽然许多基因与结肠癌的各个阶段有关，但在腺瘤形成的早期阶段发现了腺瘤性结肠息肉病（APC）基因的突变，并且认为该基因参与肿瘤的发展和进展。该基因的突变导致家族性 腺瘤性息肉病（FAP），其特征在于形成多个结肠腺瘤性息肉。这些腺瘤易于发展为结肠癌。已经开发了模拟FAP临床特征的小鼠模型（APC[Min+/o]）。因此，直接确定止血在这种癌症中的作用的研究可以在APC[Min+/o]小鼠中进行有效研究，并在止血蛋白表达方面进行额外的改变。 具体而言，本研究重点关注：1）利用自发性肿瘤模型APC[Min+/o]，止血系统组分的表达发生额外变化，并比较这些不同基因型组中的肿瘤发展和播散。这将涉及分析肿瘤生长速率、蛋白质和其他宿主细胞的空间和时间参与以及与肿瘤进展相关的血管化程度; 2)利用定量实时RT-PCR（Q-RT-PCR）鉴定和定量原发性和转移性肿瘤中血管生成因子的相对丰度，并通过原位杂交确定空间表达模式。由于我们实验室的初步研究已经证明，相对于APC [Min +/o]或APC[Min +/o]/PG-/-小鼠，APC[Min+/o]/UPA-/-小鼠中的病理学减弱。 将评估uPA/uPAR系统在肠/结肠肿瘤形成的调节中的作用。这将包括在体外和体内分析对与细胞信号传导途径相关的蛋白质的影响，以及产生具有与受体相互作用和蛋白水解活性相关的结构域特异性改变的uPA靶向突变的小鼠，以进一步阐明参与调节肿瘤形成的uPA功能结构域。

Previous studies have identified relationships between hemostasis and tumor growth and metastasis. Clinically, elevated plasma levels of these proteins have been suggested as prognostic markers for relapse and reduced survival. While these correlates exist, specific in vivo mechanisms by which this system contributes to the pathology of the disease remains largely unknown. Several clinical studies have identified components of the hemostasis system in regulating the pathology of a number of cancers. While a number of genes have been implicated in various stages of colon cancer, mutations in the adenomatous polyposis coli (APC) gene have been found during the early stages of adenoma formation, and it is believed that this gene is involved in tumor development and progression. Mutations of this gene result in familial adenomatous polyposis (FAP), which is characterized by the formation of multiple colonic adenomatous polyps. These adenomas predispose to the development of colon carcinoma. A mouse model (APC[Min+/o] )has been developed that mimics the clinical features of FAP. Therefore, studies to directly determine the role of hemostasis in this cancer can be effectively studied in APC[Min+/o] mice with additional alterations inexpression of hemostasis proteins. Specifically this study is focused on: 1) utilization of the spontaneous tumor model, APC[Min+/o], with additional alterations in expression of components of the hemostasis system and comparisons of tumor development and dissemination in these various genotypically-distinct groups. This will involve analyses of tumor growth rate, spatial and temporal involvement of proteins and other host cells, and extent of vascularization, associated with tumor progression; 2) identification and quantitation of the relative abundance of angiogenic factors in primary and metastatic tumors utilizing quantitative real-time RT-PCR (Q-RT-PCR) and determination of the spatial expression patterns by in situ hybridization. Since preliminary studies in our laboratory have demonstrated an attenuation of the pathology in APC[Min+/o]/UPA-/- mice relative to APC [Min+/o] or APC[Min+/o]/PG-/- mice 3) characterization of the mechanism associated with the uPA/uPAR system in the regulation of intestinal/colonic tumor formation will be assessed. This will include in vitro and in vivo analyses of effects on proteins associated with cell signaling pathways, and generation of mice with targeted mutations of uPA with specific alterations of domains associated with receptor interaction and proteolytic activity, in order to further elucidate functional domains of uPA involved in regulating tumor formation.