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Establishment of a nude mouse model of fluorescently tagged-colorectal cancer overexpressing hypoxia-inducible factor-1α
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Abstract:
Objective: To establish a nude mice xenograft model of fluorescently tagged-colorectal cancer overexpression of hypoxia-inducible factor-1α (HIF-1α). Methods: Four human colorectal cancer cell lines, SW480, SW620, LOVO and HCT116, were treated with CoCl2, and the one that remained HIF-1α-negative under CoCl2 induction was chosen. The selected cell line was infected with a lentiviral vector overexpressing HIF-1α and EGFP, Lenti-HIF1α-EGFP. Infected cells underwent puromycin selection. Cells stably expressing HIF-1α and EGFP. HIF-1α, confirmed by Western blotting, were used in transwell assay of migration in vitro and injected intraperitoneally into nude mice to create a xenograft model of colorectal cancer in vivo. Results: After CoCl2 induction, only SW480 remained HIF-1α-negative and was thus selected. SW480 cells stably overexpressing HIF-1α and EGFP had enhanced migration ability (250±11) as compared with control Sw480 cells (50±5) (P<0.01). The number of tumor nodules formed on the abdominal wall was significantly higher in mice injected with SW480 cells overexpressing HIF-1α (15±4) than in mice injected with control cells (4±1) (P<0.05). Conclusion: Injection of endogenously HIF-1α-negative colorectal cancer SW480 cells infected by a lentiviral vector overexpressing HIF-1α and EGFP into nude mice would provide a feasible approach of creating a xenograft model of fluorescently visible colorectal cancer for investigations on the function of HIF-1α in the pathogenesis of colorectal cancer.
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Project supported the Special Fund from Shanghai Municipal Science and Technology Commission (No.12140902302)
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