Development of in vivo experimental system for analyses of human cancer inductive microenvironment by using NOG mice

我们利用NOD/SCID/γc^（NOG）小鼠建立了一个可靠的体内肿瘤诱导微环境实验模型系统<null>。检查人胰腺癌细胞系在NOG小鼠的肝脏中形成不同转移灶的能力。利用NOG小鼠模型系统，我们建立了一个高转移细胞系，肝转移-BxPC-3（LM-BxPC-3）。我们鉴定了在LMBxPC-3细胞系中上调或下调&gt;4倍的45个基因。只有S100 A4表达与形成肝转移的能力相关，如我们在NOG小鼠中转移的定量模型中所评估的。这些结果表明，使用NOG小鼠的新实验系统为我们提供了可靠的转移模型，以寻找和开发新的抗癌疗法和针对这种和其他关键分子的新药。我们还研究了血小板反应蛋白-2（TSP-2）对人恶性黑色素瘤细胞系A375侵袭和转移的作用。我们分离了三个人恶性黑色素瘤细胞系（A375/TSP-2）转染人TSP-2。采用基质胶侵袭/迁移试验评价细胞侵袭能力。使用NOG小鼠体内实验系统进行体内转移实验，其中通过门静脉从脾脏接种肿瘤细胞。在体内转移实验中，A375/TSP-2的肝转移灶明显减少。以肝重量作为肝转移的指标，有统计学意义。组织学上也证实了表达TSP 2的转移病灶内的微血管减少。结果还表明，NOG小鼠系统可用于研究癌症诱导微环境，包括血管生成或基质蛋白酶条件。

We developed a reliable new experimental in vivo model system for assaying cancer inductive microenvironment by using NOD/SCID/γc^<null>(NOG) mice. Human pancreatic cancer cell lines were examined for their ability to form diverse metastatic foci in the livers of NOG mice. Using the NOG mouse model system, we established a highly metastatic cell line, liver metastasized-BxPC-3 (LM-BxPC-3). We identified forty-five genes that were either upregulated or down regulated >4-fold in the LMBxPC-3 cell line. Only S100A4 expression correlated with the ability to form liver metastases, as evaluated in our quantitative model of metastasis in NOG mice. These results suggested that new experimental systems using NOG mice gave us the reliable metastasis model to search for and develop new anti-cancer therapies and noveldrugs against this and other key molecules. We also examined the role of Thrombospondin-2 (TSP-2) on invasion and metastasis of human malignant melanoma cell line A375. We isolated three human malignant melanoma cell lines transfected with human TSP-2 (A375/TSP-2). Cellular invasive potential was evaluated using matrigel invasion / migration assay. The in vivo metastatic experiment was performed using the NOG mice in vivo experimental system, in which the tumor cells were inoculated via portal vein from the spleen. In the in vivo metastatic experiment, A375/TSP-2 showed obviously less liver metastatic foci. The weights of liver were measured as index for metastatic hepatomegary and disclosed statistically significant. The reduction of microvessel within metastatic lesions expressing TSP2 was also histologically confirmed. The results also demonstrated that the NOG mouse systems are useful to study cancer inductive microenvironments including angiogenesis or matrix protease conditions.