P19细胞分化过程中激活型bHLH基因的表达

目的检测P19细胞分化过程中激活型碱性螺旋-环-螺旋（bHLH）转录调控因子mRNA的表达,探讨其神经分化机制。方法用维甲酸（RA）诱导P19细胞分化,通过反转录聚合酶链反应（RT PCR）检测P19分化前以及分化后2、4?d时果蝇As c的哺乳动物类似物（Mash 1)、果蝇atonal的哺乳动物类似物(Math 1)、Neurological stem cell leukemia 2(Nscl 2) mRNA的表达。采用免疫印迹方法(Western blot)检测Mash 1蛋白的表达。结果Mash 1 mRNA和Nscl 2 mRNA 随分化表达升高,在分化后2?d到达顶峰后下降,Math 1 mRNA在分化后期表达明显。Mash 1蛋白的表达与mRNA的表达一致。结论P19细胞分化过程中bHLH转录因子mRNA的表达具有明显的时相性,为解释神经细胞分化调控机制提供实验基础。

Expression of bHLH genes in differentiation of P19 cells

To investigate the differentiation mechanisms of P19 cells by determining expression of the basic helix loop helix (bHLH) genes. MethodsP19 cells were primarily cultured and were induced to differentiate by retinoic acid(RA). Reverse transcriptase polymerase chain reaction( RT PCR) was used to detect expression of Mash 1, Math 1 and Nscl 2 mRNA in P19 cells before differentiation and at 2 and 4?d after differentiation. Western blot analysis was used to detect expression of Mash 1. ResultsExpressions of Mash 1 and Nscl 2 mRNA increased with time, and at the 2?d after differentiation they reached a peak and then decreased. Math 1 was highly expressed after differentiation. Protein expression of Mash 1 was the as same as the mRNA expression of it. ConclusionThe expression pattern of mRNA of the bHLH genes was detected, which may contribute to explaining the mechanisms of neural differentiation.