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目的:探讨雌激素对子宫内膜癌细胞系ishikawa细胞中肥胖相关基因FTO表达的调控机制以及对增殖的影响。方法:RT-PCR及细胞免疫荧光法检测不同浓度雌激素处理后ishikawa细胞中FTO表达水平的变化,PCR方法检测雌激素是否通过PI3K/AKT和MAPK信号通路对FTO进行调控,应用siRNA干扰法和MTT分析法检测FTO基因对细胞增殖的影响。结果:(1)不同浓度雌激素均可上调ishkawa细胞中FTO mRNA的表达,以10-9mol/L作用最明显,与对照组的差异有统计学意义(P<0.05);(2)E2+LY294002、E2+U0126组FTO mRNA表达水平比单独加雌激素组显著降低,差异有统计学意义(P<0.05),E2+LY294002+U0126联合加通路抑制剂组比E2+LY294002、E2+U0126单独加通路抑制剂组明显降低,差异有统计学意义(P<0.05);(3)siFTO干扰组FTO mRNA表达水平比阴性对照组明显降低,干扰效率达40%(P<0.05),FTO被干扰后,细胞的增殖活性受到明显的抑制,差异有统计学意义(P<0.05)。结论:雌激素通过受PI3K/AKT和MAPK信号通路调控的FTO基因调控细胞的增殖活性。
Objective: To investigate the regulatory mechanism of estrogen on the expression of FTO gene in ishikawa cells and its effect on proliferation. Methods: The changes of FTO expression in ishikawa cells treated with different concentrations of estrogen were detected by RT-PCR and cell immunofluorescence. Whether estrogen regulated FTO by PI3K / AKT and MAPK signal pathway was detected by PCR method. MTT assay was used to detect the effect of FTO gene on cell proliferation. Results: (1) Fetal hormones up-regulated the expression of FTO mRNA in ishkawa cells with different concentrations of estrogen, with the most obvious effect at 10-9 mol / L, which was significantly different from the control group (P <0.05); (2) The levels of FTO mRNA in LY294002 and E2 + U0126 groups were significantly lower than those in estrogen alone group (P <0.05), and those in E2 + LY294002 + U0126 group plus E2 group were higher than those in E2 + LY294002 and E2 + U0126 groups (3) The expression level of FTO mRNA in siFTO interference group was significantly lower than that in negative control group, the interference efficiency was 40% (P <0.05), FTO was disturbed After the cell proliferation activity was significantly inhibited, the difference was statistically significant (P <0.05). CONCLUSIONS: Estrogen regulates the proliferation of cells via the FTO gene regulated by the PI3K / AKT and MAPK signaling pathways.