DOI: 10.46810/tdfd.1770061 ISSN: 2149-6366

Detection of Micrometastases of Luciferase-Expressing LLC-1 Lung Cancer Cells in a Mouse Metastasis Model: Comparison of qPCR and Bioluminescence Imaging

Süleyman Can Öztürk
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This study investigates the detection of micrometastases in a mouse model of lung cancer using luciferase-expressing Lewis Lung Carcinoma cells (LLC1-Luc2), comparing bioluminescence imaging and quantitative PCR targeting the sex determining region Y gene. LLC1-Luc2 cells were intravenously injected into female C57BL/6 mice to establish a syngeneic metastasis model. Metastatic progression in lungs and liver was monitored over four weeks using bioluminescence imaging, with luciferase signals recorded on days 1, 7, 14, 21, and 28. Concurrently, quantitative PCR was employed to quantify metastatic cell numbers by amplifying the male-specific sex determining region Y gene, leveraging the sex mismatch between male-derived LLC1-Luc2 cells and female hosts. A standard curve was generated to correlate quantitative PCR cycle threshold values with cell numbers, enabling precise estimation of metastatic burden. Results demonstrated that BLI effectively visualized metastatic foci in deep tissues, while quantitative PCR provided superior sensitivity for detecting micrometastases, particularly at early stages or in low-cell-number lesions. The integration of bioluminescence imaging and sex determining region Y gene quantitative PCR offers a robust framework for both qualitative and quantitative assessment of metastasis, providing valuable insights for preclinical evaluation of anti-metastatic therapies

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