Hemostasis System in Tumor Growth and Metastasis

以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，这些蛋白的血浆水平升高被认为是复发和生存率下降的预后标志。虽然存在这些相关性，但该系统在疾病病理中的具体作用机制在很大程度上仍不清楚。几项临床研究已经确定了止血系统在调节一些癌症的病理过程中的组成部分。虽然许多基因与结肠癌的不同阶段有关，但在腺瘤形成的早期阶段就发现了腺瘤性息肉病结肠(APC)基因的突变，人们认为该基因参与了肿瘤的发生和发展。该基因的突变导致家族性 腺瘤性息肉病(FAP)，以多发性结肠腺瘤性息肉的形成为特征。这些腺瘤易发生结肠癌。已经建立了一种模拟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]小鼠身上进行有效的研究，同时止血蛋白的表达也发生了额外的变化。 这项研究的重点是：1)利用自发性肿瘤模型APC[Min+/o]，进一步改变止血系统成分的表达，并比较这些不同基因型别人群中肿瘤的发展和扩散情况。这将涉及与肿瘤进展相关的肿瘤生长速度、蛋白质和其他宿主细胞的空间和时间受累以及血管形成程度的分析； 2)用实时定量RT-PCR方法鉴定和定量原发和转移瘤中血管生成因子的相对丰度，并用原位杂交法确定其空间表达模式。由于我们实验室的初步研究表明，与APC[Min+/o]或APC[Min+/o]/Pg-/-小鼠相比，APC[Min+/o]/uPA-/-小鼠的病理学减弱3)与 将评估uPA/uPAR系统在调节肠/结肠肿瘤形成中的作用。这将包括体外和体内对与细胞信号通路相关的蛋白质的影响的分析，以及通过与受体相互作用和蛋白分解活性相关的结构域的特定变化的uPA靶向突变的小鼠的产生，以进一步阐明参与调节肿瘤形成的uPA的功能结构域。

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.