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Sensitive optical detection of an early metastatic tumor using a new cell line with enhanced luminescent and fluorescent signals

Keywords: dual-labeling , luciferase activity , GFP , prostate cancer cells , metastasis , optical imaging

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Abstract:

Animal models using cell lines that are dual-labeled with luciferase and green fluorescent protein (GFP) are powerful tools for performing simultaneous quantitative and qualitative analysis of metastatic tumors. However, the applications of such dual-labeled tumor models have been limited due to the technical challenges associated with low bioluminescent signaling from tumor cells. Here, we used lentiviral vector (LV) encoding firefly luciferase and GFP and engineered a more sensitive and highly metastatic prostate cancer cell line (MLL-Luc/GFP cells), which allows simultaneous fluorescence and luminescence imaging. The light emission of MLL-Luc/GFP cells was 33.5 fold higher than that of PC-3-luc2-GFP prostate cancer cells which showed 750 p/s light emission per cell. Furthermore, the MLL-Luc/GFP cells showed 3.9 fold higher luciferase activities than did 4T1-luc2, which was previously recognized as exhibiting the highest luciferase activity. An in vivo evaluation with optical imaging showed pinpoint localization of GFP-positive cells in a metastatic lung as well as easy detection of early metastatic spreading. The newly engineered MLL-Luc/GFP cells provide an appropriate metastatic animal model system for future studies of metastasis and the testing of anti-metastatic therapies specifically aimed at prostatic cancer.

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