目的 建立稳定表达FOXO1-GFP的人骨肉瘤U2OS细胞系，并验证U2OS-FOXO1-GFP细胞模型对氧化应激损伤的指示作用。方法 将pcDNA3-GFP-FOXO1质粒转染到U2OS细胞中，利用G418和FCM筛选稳定细胞系。40mmol/L AAPH处理U2OS-FOXO1-GFP 细胞4 h，CCK-8检测细胞活力，荧光显微镜观察FOXO1-GFP的分布情况。结果 成功获得U2OS-FOXO1-GFP稳转细胞系。经G418和FCM筛选后，FOXO1-GFP阳性细胞占细胞总数的92%。CCK-8结果显示，AAPH处理后，U2OS-FOXO1-GFP细胞存活率下降至0.55±0.04明显低于对照组（P＜0.05）。荧光显微镜观察发现，正常情况下，FOXO1-GFP位于细胞质；AAPH处理后，FOXO1-GFP易位到细胞核。结论 U2OS-FOXO1-GFP细胞系能很好的指示细胞氧化应激损伤，为筛选抗氧化应激药物提供了一种有利工具。

Abstract: Objective To establish the stable FOXO1-GFP transfected U2OS cell line and detect U2OS-FOXO1-GFP indicative function of oxidative stress. Methods The FOXO1-GFP vector was transfected into U2OS cell. Stably transfected U2OS cell was screened using G418 and Flow Cytometer. After 4 hours treatment of AAPH, cell survival was tested by CCK-8 and the location of FOXO1-GFP was observed by fluorescence microscope. Results The stable FOXO1-GFP transfected U2OS cell line was successfully established. The percentage of FOXO1-GFP positive cells in total cells was 92% after screened through G418 and FCM. CCK-8 assay showed that when U2OS-FOXO1-GFP cells were treated with AAPH, cell survival dropped to 0.55±0.04 compared with control (P＜0.05). In control vehicle group, FOXO1-GFP was localized in the cytosol was observed by fluorescence microscope. When cells were treated with 40 mmol/L AAPH 4 hours, translocation of FOXO1-GFP to the nucleus was observed. Conclusion The U2OS-FOXO1-GFP cell line can indicate the oxidative stress. It would be a convenient tool to study the antioxidant responses drugs.