Dooley T P, Stamp-Cole M, Ouding R J
Department of Genetics, Southwest Foundation for Biomedical Research, San Antonio, TX 78228.
Lab Anim Sci. 1993 Feb;43(1):48-57.
We developed and evaluated an in vivo athymic nude mouse model for tumor growth, angiogenesis, metastasis, and antineoplastic drug development. Melanoma cell lines expressing beta-galactosidase encoded by the Escherichia coli lac Z gene have been created by infecting an immortal murine melanocyte cell line with a recombinant retrovirus expressing the v-Ha-ras oncogene and lac Z to generate the MRB (melanoma, ras, beta-galactosidase) cell lines. The amelanotic, phorbol ester-independent, transformed melanoma cell lines developed tumors rapidly when injected subcutaneously into nude mice, as well as experimental lung metastases when injected i.v. into the tail vein. beta-galactosidase-expressing subcutaneous tumors and lung metastases stained blue with X-gal. The melanomas produced in nude mice have been characterized by using various histochemical and immunohistochemical staining methods to detect melanoma- and endothelial-cell-specific markers to determine the extent of neovascularization in MRB nude mouse tumors. Optimal staining of endothelial cells involved in tumor angiogenesis was observed by using ADPase activity and antiangiotensin-converting enzyme antibody staining. Attempts at indirect quantification of metastatic tumor cell number within the lung by either beta-galactosidase enzymatic activity or ELISA immunoreactivity were unsuccessful. However, the MRB cell lines should be useful in screening for and studying the mechanisms of action of antineoplastic, antimetastatic, and angiostatic drugs in vivo in athymic nude mice.
我们开发并评估了一种用于肿瘤生长、血管生成、转移及抗肿瘤药物研发的无胸腺裸鼠体内模型。通过用表达v-Ha-ras癌基因和lac Z的重组逆转录病毒感染永生小鼠黑素细胞系，已构建出表达大肠杆菌lac Z基因编码的β-半乳糖苷酶的黑色素瘤细胞系，以产生MRB（黑色素瘤、ras、β-半乳糖苷酶）细胞系。无黑色素、佛波酯非依赖性的转化黑色素瘤细胞系皮下注射到裸鼠体内时能迅速形成肿瘤，静脉注射到尾静脉时会产生实验性肺转移。表达β-半乳糖苷酶的皮下肿瘤和肺转移灶用X-gal染色呈蓝色。已通过使用各种组织化学和免疫组织化学染色方法对裸鼠体内产生的黑色素瘤进行了表征，以检测黑色素瘤和内皮细胞特异性标志物，从而确定MRB裸鼠肿瘤中的新血管生成程度。通过使用ADP酶活性和抗血管紧张素转换酶抗体染色观察到参与肿瘤血管生成内皮细胞的最佳染色效果。通过β-半乳糖苷酶酶活性或ELISA免疫反应性间接定量肺内转移瘤细胞数量的尝试未成功。然而，MRB细胞系在筛选和研究无胸腺裸鼠体内抗肿瘤、抗转移和血管生成抑制药物的作用机制方面应是有用的。
