ERα特异性抑制剂MPP降低小鼠2-细胞胚G1/S期过渡相关因子Nanog、cyclin D1和cyclin E水平

Estrogen receptor α specific antagonist MPP inhibits G1/S transitionrelated factors Nanog, cyclin D1 and cyclin E in mouse 2-cell embryos

目的观察雌激素受体(ERa)特异性抑制剂甲基哌啶吡唑(methyl-piperidino-pyrazole,MPP)对小鼠2-细胞胚G1/S期过渡相关因子Nanog、cyclin D1和cyclin E表达水平的影响,阐明ERa对小鼠2-细胞胚G1/S期过渡的作用,为进一步探讨ERa在小鼠植入前胚合子基因组激活中的作用机制提供理论依据。方法收集昆明雌性小鼠与雄性小鼠交配所得的受精卵,分别置于无MPP的培养液和含20mmol/L MPP的培养液中培养,获取体外发育的2-细胞胚,采用免疫荧光染色技术检测细胞内Nanog、cyclin D1和cyclin E水平。结果免疫荧光染色结果显示,含20mmol/L MPP培养液培养的受精卵其体外发育2-细胞胚内Nanog、cyclin D1和cyclin E免疫反应性明显降低。结论 ERa可通过调控小鼠2-细胞胚内Nanog、cyclin D1和cyclin E水平,影响细胞从G1期到S期的过渡。

Objective To investigate the effect of estrogen receptorα(ERα)specific antagonist methyl-piperidino-pyrazole(MPP)on the expression of G1/S transition-related factors Nanog,cyclin D1and cyclin E in mouse2-cell embryos,so as to demonstrate the role of ERαin G1/S transition in mouse2-cell embryos and provide a theoretical basis for further research on the mechanism of the activation of pre-implantation mouse zygotic genome by ERα.Methods Zygotes were collected from Kunming female mice that had been mated with male mice,and cultured in media without MPP and media supplemented with20mol/L MPP,respectively.The2-cell embryos were obtained,and the levels of Nanog,cyclin D1and cyclin E were detected by immunofluorescent staining.Results Immunofluorescent staining results showed that the immunoreactivity of Nanog,cyclin D1and cyclin E in the2-cell embryos cultured in20mol/L MPP was obviously lower than in those cultured without MPP treatment.Conclusion ERαmight affect G1/S transition via regulating the expression of Nanog,cyclin D1and cyclin E in mouse2-cell embryos.