MODEL TO SEQUENTIALLY ANALYZE THE METASTATIC CASCADE

实体瘤在人和动物中的转移性扩散需要 已成功完成连续步骤。除了增长之外， 原发灶，细胞必须进入血管内，存活的运输过程中 可能需要横切淋巴结，在靶器官等 如肺或肝，在目标的实质内渗出/建立 并复制形成临床上有意义的转移性结节。 我们建议开发的模型将由小鼠的亚群组成 乳腺肿瘤具有可选择的抗药性标记，使我们能够 以监测转移扩散的动力学和途径。这个 酶促培养后形成集落的肿瘤细胞的恢复 选择性介质中分散的组织提供了一种高度敏感的方法 以定量的方式检测隐匿性转移。通过将一种 视觉标记进入细胞(E.ColiB-半乳糖苷酶基因)，休眠 也可检测到非克隆性细胞。 一组通过不同途径转移的转移亚群 (即血液和淋巴途径)至不同部位(即肝脏) 和/或肺)和在不同阶段失败的非转移亚群 在转移序列中是要被推导和表征的。几个 已经获得了变种，并对其进行了部分表征， 说明了这种方法的潜力。这些不同的留置权是 在细胞培养中易于维护，易于分类和回收 液氮是高度致癌的，并且对 致癌、转移和药物标记物特性。 这一模型最终将使人们能够准确地确定特定的 肿瘤进展特定阶段的宿主免疫机制 在转移过程中扩散。

Metastatic dissemination of solid tumors in man and animals requires the successful completion of sequential steps. In addition to growth of the primary lesion, cells must intravasate, survive the transport process which may require the transversal of lymph nodes, arrest in a target organ such as the lung or liver, extravasate/establish in the parenchyma of the target organ, and replicate to form clinically significant metastatic nodules. the model we propose to develop will consist of subpopulations of a mouse mammary tumor which have selectable drug resistance markers that allow us to monitor the kinetics and route of metastatic dissemination. The recovery of colony forming tumor cells after plating enzymatically dispersed tissues in selective media provides a highly sensitive method to detect occult metastases in a quantitative manner. By transfecting a visual marker into the cells (E.coli B-galactosidase gene), dormant nonclonogenic cells may also be detected. A panel of metastatic subpopulations which metastasize via different routes (i.e. hematogenous and lymphatic routes) to different sites (i.e. liver and/or lung) and nonmetastatic subpopulations which fail at different steps in the metastatic sequence are to be derived and characterized. Several variants have already been obtained and partially characterized, illustrating the potential of this approach. These variant liens are easily maintained in cell culture, are readily sorted and recovered from liquid nitrogen, are highly tumorigenic, and are stable with respect to tumorigenic, metastatic, and drug marker properties. This model will ultimately allow one to pinpoint the effect of specific host immune mechanisms on specific stages of tumor progression and dissemination during metastasis.