目的：建立一种荧光可视化显像且稳定过表达缺氧诱导因子-1α（hypoxia-inducible factor-1α，HIF-1α）的裸鼠结直肠癌移植瘤模型。方法：对不同的人结直癌细胞系（SW480、SW620、LOVO及HCT116细胞）用缺氧诱导剂CoCl2处理，根据HIF-1α被诱导表达的情况选择合适的靶细胞；将HIF-1α的cDNA序列克隆至慢病毒表达质粒pLV-TRC-EGFP，构建pLV-HIF1α-EGFP慢病毒表达质粒，并包装过表达HIF-1α的慢病毒颗粒Lenti-HIF1α-EGFP。将Lenti-HIF1α-EGFP感染SW480细胞，嘌呤霉素筛选稳定过表达HIF-1α的细胞株SW480HIF-1α，Western blotting检测SW480HIF-1α细胞内HIF-1α及其下游蛋白VEGF、M1型丙酮酸激酶（pyruvate kinase expression M1，PKM1）的表达情况，Transwell实验检测其迁移能力。裸鼠腹腔注射SW480HIF-1α建立结直肠癌移植瘤模型，倒置荧光显微镜观察裸鼠腹腔移植瘤结节。结果：常氧条件下4种结直肠癌细胞均不表达HIF-1α，经CoCl2诱导后，除SW480细胞系以外的细胞均可被诱导表达HIF-1α，故选择SW480细胞作为研究靶细胞。成功构建稳定过表达HIF-1α的细胞株SW480HIF-1α，SW480HIF-1α细胞内HIF-1α、VEGF和PKM1的表达水平均高于野生型SW480细胞，其迁移能力较野生型SW480细胞显著增强\[观察视野内迁移细胞数：（250±11）vs（50±5）个，P<0.01\]。SW480HIF-1α组裸鼠肠壁形成肿瘤结节数量显著多于SW480组\[（15±4）vs（4±1）个，P<0.05\]。结论：成功建立了高表达HIF-1α的荧光可视化裸鼠移植瘤模型，为进一步的相关功能学研究以及药物筛选提供条件。

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 CoCl2, 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.

上海市科学技术委员会专项基金资助项目（No.12140902302）。