黑色素瘤小鼠高转移模型体内筛选和细胞系的建立  

Establishment of a Highly Metastatic Model of Melanoma in Mice in vivo Selection and Cell Line

作　　者：吴海燕[1] 姚明[2] 闫明霞[2] 刘蕾[2] 孔韩卫[2] 薛整风[1] 

机构地区：[1]扬州大学兽医学院,扬州225009 [2]上海市肿瘤研究所,上海200032

出　　处：《实验动物与比较医学》2008年第2期67-73,共7页Laboratory Animal and Comparative Medicine

基　　金：上海市科委登山行动计划(064909001)

摘　　要：目的采用体内综合筛选的方法建立B16黑色素瘤小鼠高转移模型和相应细胞系,为系统性研究肿瘤转移机制和临床肿瘤治疗提供动物模型。方法首先将小鼠黑色素瘤B16移植于T、B、NK免疫细胞缺陷的BNX小鼠皮下,通过皮下移植→肺转移→皮下移植→肺转移的体内循环筛选方法筛选。在前三代筛选过程中,肿瘤直径达1.5 cm时切除皮下瘤,延长小鼠寿命以获得明显肺转移灶。在后三代体内筛选过程中,待小鼠自然濒死时,剖解获得肺转移灶。共通过体内筛选的方法筛选六代,建立高转移细胞系。同时进行肿瘤的生长和转移情况观察、病理组织学、细胞生长曲线测定、流式细胞术分析、染色体分析和细胞侵袭基底膜实验。结果BNX小鼠切瘤筛选3代后肺转移率达80%,转移程度达+++,转移天数平均达45 d。直接皮下筛选第一代转移程度和转移率有所下降,但三代后,转移率也达到91.7%(11/12),转移程度也达到+++,转移天数缩短至35 d。病理组织学、细胞生长曲线、流式细胞术和染色体分析等结果表明与黑色素瘤生物学特性相似。结论本研究建立了一个肺转移率高、转移特性稳定、直观性好、操作简便的小鼠黑色素瘤B16自发高转移模型及相应的细胞系,为探讨肿瘤转移的生物学机制和抗转移治疗提供了理想的动物模型。Objective To establish a highly metastatic model and cell line of melanoma B 16 in mice by using organ selection of metastasis. Methods Histologically melanoma B16 intact tissues were subcutaneously implanted into the mice. In order to gain visible metastatic focus, BNX mice with immunodeficiency was selected according to the method of subcutaneous implantation → lung metastasis → subcutaneous inoculation → lung metastasis. In the first three generations neoplasma were removed by tumor resection when it reached 1.5 cm in diameter. In the following three generations the same steps were duplicated without resection. Then homologous cell line was established. Tumorgenicity, invasion, metastasis and morphological characteristics of the implanted tumors were studied by light microscopy, histology, cell growth curve, flow cytometry, Chromosome analysis and basement membrane invasion assay. Results Highly metastatic models of melanoma B 16 were obtained after organ screening. After 3 generations in vivo, the carcinoma metastasis rate accounted to 80 percent, metastasis extent reached ＋＋＋ after 45 days. While only a lower metastasis rate was found when the tumor was subcutaneously implanted absolutely without resection at the first time. But a 91,7 percent lung metastatic rate and high metastasis extent were observed again after being passaged in vivo for 3 generations after 35 days per generation. It indicated that 1316-sci has similar characteristic with 1316 by the assay about histology, cell growth curve, flow cytometry, Chromosome analysis and basement membrane invasion. Conclusions we had established a highly metastasis melanoma 1316 model and homologous cell line, which was high, steady, and intuitionistic metastasis and have same biological characteristics of mice melanoma B 16. The result provided a useful tool for the study of metastatic mechanism and treatment of carcinoma.

关 键 词：黑色素瘤 BNX小鼠 筛选 转移 动物模型 

分 类 号：R739.5[医药卫生—肿瘤]