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目的比较三种细胞在建立脆性X综合征细胞模型中的差异,为脆性X综合征的实验研究提供有效,便利的细胞模型。方法利用硝普钠可通过产生NO诱导脆性X智力低下一号基因(fragile X mental retardation-1 gene,FMR1)启动子区Cp G岛甲基化的原理,建立脆性X综合征细胞模型;采用人类新鲜的外周血单个核细胞(PMBC),大鼠肾上腺嗜铬细胞瘤细胞(PC12),人类慢性髓源性白血病细胞系(K-562),比较三种细胞模型,在硝普钠用药浓度,封闭持续时间及用腺苷酸环化酶激活剂弗司可林(forskolin,FSK)诱导封闭基因重新表达持续时间等方面的特点,通过逆转录PCR检测FMR1基因封闭或开启的效果。结果三种细胞模型在硝普钠用药浓度,封闭持续时间及弗斯可林诱导重新表达持续的时间方面都不完全相同。结论三种脆性X综合征细胞模型各有其优缺点,可根据不同地研究方向选用不同的细胞模型。
OBJECTIVE: To compare the differences of three kinds of cells in establishing a cell model of fragile X syndrome and to provide an effective and convenient cell model for the experimental study of Fragile X syndrome. Methods Using nitroprusside sodium could induce fragile X mental retardation-1 gene (FMR1) promoter CpG island methylation in the principle of fragile X syndrome cell model; using human Fresh peripheral blood mononuclear cells (PMBC), rat adrenal pheochromocytoma cells (PC12) and human chronic myelogenous leukemia cell line (K-562) were compared in three cell models in the concentration of sodium nitroprusside, The duration of blocking, and the duration of re-expression of the enclosed gene induced by the adenylate cyclase activator forskolin (FSK). The effect of blocking or opening the FMR1 gene was examined by reverse transcription PCR. Results The three cell models were not identical in the concentration of sodium nitroprusside, the duration of occlusion and the duration of forskolin-induced reexpression. Conclusion The three kinds of fragile X syndrome cell models have their own advantages and disadvantages. Different cell models can be selected according to different research directions.