Abstract
BackgroundBioluminescent imaging (BLI) is based on the detection of light emitted by living cells expressing a luciferase gene. Stable transfection of luciferase in cancer cells and their inoculation into permissive animals allows the noninvasive monitorization of tumor progression inside internal organs. We have applied this technology for the development of a murine model of colorectal cancer involving the liver, with the aim of improving the pre-clinical evaluation of new anticancer therapies.ResultsA murine colon cancer cell line stably transfected with the luciferase gene (MC38Luc1) retains tumorigenicity in immunocompetent C57BL/6 animals. Intrahepatic inoculation of MC38Luc1 causes progressive liver infiltration that can be monitored by BLI. Compared with ultrasonography (US), BLI is more sensitive, but accurate estimation of tumor mass is impaired in advanced stages. We applied BLI to evaluate the efficacy of an immunogene therapy approach based on the liver-specific expression of the proinflammatory cytokine interleukin-12 (IL-12). Individualized quantification of light emission was able to determine the extent and duration of antitumor responses and to predict long-term disease-free survival.ConclusionWe show that BLI is a rapid, convenient and safe technique for the individual monitorization of tumor progression in the liver. Evaluation of experimental treatments with complex mechanisms of action such as immunotherapy is possible using this technology.
Highlights
Bioluminescent imaging (BLI) is based on the detection of light emitted by living cells expressing a luciferase gene
Characterization of MC38 colon cancer cells stably transfected with the luciferase gene MC38 cells were transfected with a plasmid carrying luciferase under the control of the CMV promoter, and 10 different clones were isolated after antibiotic selection
We represent light emission in living cells corresponding to 3 clones after incubation of increasing number of cells with the substrate D-luciferin
Summary
Bioluminescent imaging (BLI) is based on the detection of light emitted by living cells expressing a luciferase gene. Stable transfection of luciferase in cancer cells and their inoculation into permissive animals allows the noninvasive monitorization of tumor progression inside internal organs. We have applied this technology for the development of a murine model of colorectal cancer involving the liver, with the aim of improving the pre-clinical evaluation of new anticancer therapies. 10–25% of colon cancer patients present one or multiple liver metastases at the time of diagnose [1]. Hepatic metastases from colon cancer are frequently observed in the clinic and they are the most frequent cause of death in these patients. Advances in the management of this disease will probably require the combination of standard care and new therapies that are still in the experimental stage
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