缺氧对HCT116肠癌细胞成纤维细胞生长因子21的调节作用

Regulation of hypoxia on fibroblast growth factor 21 in HCT116 intestinal cancer cells

目的观察缺氧模拟物二氯化钴(CoCl2)对人源HCT116肠癌细胞成纤维细胞生长因子21(FGF21)表达的影响。方法分别用0、50、100和200μmol/L的CoCl2处理HCT116细胞24 h,同时用200μmol/L的CoCl2处理HCT116细胞12、48 h,RT-PCR法和Western blot法分别检测细胞中的FGF21 mRNA和蛋白的表达水平;分别用缺氧诱导因子抑制剂2ME2和YC-1,同时加入CoCl2处理HCT116肠癌细胞24 h,RT-PCR法检测细胞中的FGF21 mRNA;用抗氧化剂NAC和CoCl2同时处理HCT116肠癌细胞24 h,RT-PCR法和Western blot法分别检测细胞中的FGF21 mRNA和蛋白的表达量。结果随CoCl2浓度增加和作用时间延长,HCT116细胞FGF21mRNA相对表达量逐渐降低(P<0.05)。Western blot结果显示,与对照组相比,CoCl2组FGF21蛋白表达水平显著下降(P<0.05)。缺氧诱导因子抑制剂处理后,HCT116细胞FGF21 mRNA相对表达量依然降低(P<0.05)。抗氧化剂处理后,HCT116细胞FGF21 mRNA和蛋白相对表达量未出现明显降低(P>0.05)。结论CoCl2对HCT116肠癌细胞FGF21的表达有抑制作用,该作用与缺氧诱导因子无关,而与氧化应激有关。

Objective To observe the effect of hypoxia mimicking cobalt dichloride(CoCl2)on the expression of fibroblast growth factor 21(FGF21)in human HCT116 intestinal cancer cells.Methods HCT116 cells were treated with 0,50,100 and 200μmol/L CoCl2 for 24 h,and HCT116 cells were treated with 200μmol/L CoCl2 for 12 and 48 h.RT-PCR and Western blot were used to detect the expression of FGF21 mRNA and protein in the cells.HCT116 intestinal cancer cells were treated with hypoxia-inducible factor inhibitors 2ME2 and YC-1,respectively,added into CoCl2 for 24 h.RT-PCR was used to detect FGF21 mRNA in cells.HCT116 intestinal cancer cells were treated with antioxidants NAC and CoCl2 for 24 h,and the expression of FGF21 mRNA and protein was detected by RT-PCR and Western blot.Results With the increase of CoCl2 concentration and prolonged action time,the relative expression of FGF21 mRNA in HCT116 cells decreased gradually(P<0.05).Western blot results showed that the expression of FGF21 protein in CoCl2 group was significantly lower than that in the control group(P<0.05).After treatment with hypoxia-inducible factor inhibitor,the relative expression of FGF21 mRNA in HCT116 cells was still decreased(P<0.05).After antioxidant treatment,the relative expression of FGF21 mRNA and protein in HCT116 cells did not decrease significantly(P>0.05).Conclusion CoCl2 can inhibit the expression of FGF21 in HCT116 intestinal cancer cells,which is not related to hypoxia-inducible factors but related to oxidative stress.