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通过脂质体介导 gfp表达质粒转移及G418筛选获得了稳定表达 gfp的小鼠膀胱移行细胞癌细胞株BTT GFP。利用 gfp作为肿瘤细胞的标记 ,结合罗丹明标记的葡聚糖尾静脉注射作血管造影 ,建立了一种新的肿瘤模型 ,具有简便、可靠、无创伤的优点 ,特别是可以通过荧光显微镜动态观察肿瘤转移病灶形成最早期阶段肿瘤细胞生长与肿瘤局部宿主血管的变化。利用新型鼠耳肿瘤模型观察到移植的肿瘤细胞会主动向宿主血管迁移 ,当肿瘤生长至仅 0 .3mm直径大小时即可见血管生成。免疫组织化学染色观察到肿瘤内新生血管SMA及CD31染色阳性 ;肿瘤细胞不仅高水平表达VEGF ,也高水平表达VEGF的受体Flk 1。提示肿瘤局部存在VEGF自分泌与旁分泌通路 ,肿瘤细胞高水平表达VEGF是新生血管生成的主要原因
The gfp mouse bladder transitional cell carcinoma cell line BTT GFP stably expressing gfp was obtained by liposome-mediated gfp expression plasmid transfer and G418 screening. Using gfp as a tumor cell marker and combining with rhodamine labeled dextran intravenous injection for angiography, a new tumor model has been established, which has the advantages of simplicity, reliability and non-invasiveness, and can be observed by fluorescence microscopy in particular Tumor metastasis in the earliest stages of tumor cell growth and tumor local host vessel changes. Transplanted tumor cells were observed to migrate actively to the host vessel using the novel mouse ear tumor model and angiogenesis was seen when the tumor grew to a size of only 0.3 mm diameter. Immunohistochemical staining showed that there were positive staining of SMA and CD31 in the neoplasm of the tumor. The tumor cells not only expressed VEGF at a high level but also expressed VEGF receptor Flk 1 at a high level. Suggesting that there is VEGF autocrine and paracrine pathways in tumor, and the high level of VEGF expression in tumor cells is the main reason of neovascularization