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c-Fos对结直肠癌细胞HCT-116影响

Effect of c-Fos on proliferation and apoptosis in colorectal cells HCT116

  • 摘要: 目的 探讨AP-1转录因子组成成分之一c-Fos对结直肠癌细胞增殖和凋亡的影响。方法 将合成的siRNA转染入HCT-116细胞中,实时定量PCR法检测其中c-Fos的含量;利用四甲基偶氮唑盐(MTT)显色法检测c-Fos被敲低后对细胞生长增殖的影响;Etoposide诱导细胞凋亡,溴化丙啶(PI)染色、流式细胞仪检测细胞凋亡的改变。结果 siRNA转染结直肠癌细胞HCT-116 48 h后,与对照组相比,c-Fos的含量降低约50%;MTT实验中RNAi组细胞增殖速度时间增加,加入etoposide(100 μmol/L)12 h后,对照组细胞凋亡率为(27.6±2.07)%,RNAi组细胞的凋亡率为(39.1±4.84)%,凋亡率降低。结论 c-Fos在HCT-116细胞中发挥着促进细胞生长和抑制凋亡的作用。

     

    Abstract: Objective To explore the effect of c-Fos on proliferation and apoptosis in colorectal cancer cells HCT-116.Methods C-Fos siRNA was transfected into HCT-116 cell and quantitative real-time PCR was used to measure relative expression of c-Fos.The difference in cell proliferation between control group and RNAi group was detected with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay.Propidium iodide(PI)staining and fluorescence-activated cell sorter(FACS)assay were adopted to measure apoptosis in c-Fos knockdown cells after etoposide treatment.Results The relative expression of c-Fos in HCT-116 cells was decreased by about 50% 48 hours after c-Fos siRNA was transfected into the cells.The results of MTT assay indicated that the cell growth of RNAi group was significantly accelerated at 1,2,3,4,5 days,with a time-dependent manner.Twelve hours after the cell apoposis was induced by etoposide(100 μM),the percentage of apoptosis in control group and RNAi group was 27.6±2.07 and 39.1±4.84%,respectively.Conclusion C-Fos could promote cell proliferation and inhibit apoptosis in colorectal cancer cells HCT-116.

     

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