A model of spontaneous lung metastases visualised in fresh host tissue by green fluorescent protein expression

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.