Paris S, Chauzy C, Martin-Vandelet N, Delpech B, Thiberville L, Martin J P, Diarra-Mehrpour M
INSERM U295, Faculté de Médecine, Rouen, France.
Clin Exp Metastasis. 1999;17(10):817-22. doi: 10.1023/a:1006782307004.
The authors describe a model of spontaneous lung metastases in nude mice using green fluorescent protein (GFP) expression as a marker. The human lung cell line H460M was transfected with the humanised GFP-S65T cDNA and a stable fluorescent cell line termed H460M(GFP) was obtained. The latter kept in vitro biological features when compared to the parental H460M cell line, which suggests that GFP-expression does not influence H460M(GFP) cell line behaviour. In order to evaluate their metastatic potential and to determine the number of spontaneous metastases, H460M(GFP) cells were subcutaneously inoculated into nude mice. Animals were sacrificed at time intervals and tissues (lung, liver, spleen, node, and kidney) were analysed under fluorescence microscopy. These experiments demonstrated that 2 weeks after subcutaneous inoculation, 75% of animals exhibited fluorescent spontaneous lung micrometastases. From the third week, 100% of animals exhibited an increasing number of metastases (10-16) which were only localised in the lungs. At the end of the study, the number of lung metastases had dramatically increased (42-400 at 7 weeks). Although these metastases were mainly localised in lung, a few mice had an invasion of neighbouring lymph nodes. The H460M(GFP) cell line allowed to follow the seeding and development of spontaneous lung metastases and may be considered a simple and powerful tool to study each step of the metastasis to screen new anticancer drugs.
作者描述了一种在裸鼠中使用绿色荧光蛋白（GFP）表达作为标记的自发性肺转移模型。将人肺细胞系H460M用人类化的GFP-S65T cDNA转染，获得了一个稳定的荧光细胞系，称为H460M(GFP)。与亲本H460M细胞系相比，后者保留了体外生物学特性，这表明GFP表达不影响H460M(GFP)细胞系的行为。为了评估它们的转移潜能并确定自发性转移的数量，将H460M(GFP)细胞皮下接种到裸鼠体内。在不同时间间隔处死动物，并在荧光显微镜下分析组织（肺、肝、脾、淋巴结和肾）。这些实验表明，皮下接种2周后，75%的动物出现了荧光性自发性肺微转移。从第三周开始，100%的动物转移灶数量增加（10-16个），且仅局限于肺部。在研究结束时，肺转移灶数量急剧增加（7周时为42-400个）。虽然这些转移灶主要局限于肺部，但少数小鼠出现了邻近淋巴结的侵袭。H460M(GFP)细胞系能够追踪自发性肺转移的播种和发展，可被视为研究转移各个步骤以筛选新抗癌药物的一种简单而强大的工具。
