MECHANISMS AND INHITION OF THE INVASION OF MALIGNANT BONE AND SOFT TISSUE TUMORS THROUGH BASEMENT MEMBRANES

肿瘤对基底膜的侵袭是导致转移形成的复杂的多阶段过程中的关键一步。肿瘤细胞在扩散过程中最初侵入血管床时会穿过基底膜，然后穿透目标组织。人纤维肉瘤细胞HT1080经cAMP类似物和梭鱼毒素处理后，细胞内cAMP浓度升高，TIMP-1和TIMP-2在mRNA和蛋白水平均有表达。这种TIMPs的诱导与HT1080细胞的IV型胶原溶解活性、侵袭力和转移潜能有很好的相关性。我们建立了一种使用基底膜提取物(Matrigel)来定量评估体内肿瘤血管生成的方法。裸鼠皮下注射。将液体Matrigel与HT1080细胞混合。由于Matrigel在体温下迅速形成固体凝胶，含有肿瘤细胞的凝胶可以立即被去除，并加工用于组织学研究。通过使用图像分析仪测量凝胶中存在的新生血管的数量和总面积来定量监测肿瘤血管生成。

Tumor invasion of the basement membranes is a crucial step in the complex multistage process which leads to the formation of metastasis. Tumor cells cross the basement membranes as they initially invade vascular beds during dissemination, and then they penetrate their target tissue. The treatment of human fibrosarcoma cells HT1080 with cAMP analogues and chorela toxin that increase the intra-cellular cAMP concentrations induces the expression of Timp-1 and Timp-2 both at the mRNA and the protein levels. This induction of Timps correlated well with type IV collagenolytic activity, invasiveness and metastatic potential of HT1080 cells.We developed a quantitative assay for assessing in vivo tumor angiogenesis using basement membrane extracts (Matrigel). Nude mice are injected s.c. with liquid Matrigel mixed with HT1080 cells. Since Matrigel forms a solid gel rapidly at body temperature, the gel containing tumor cells can be removed immediately, and the processed for histological studies. Tumor angiogenesis was monitored quantitatively by measuring both the number and total area of neovessels present in the gels using an image analyzer.