PMA诱导C6细胞rPer1,rDbp基因的近日节律表达

目的寻找一种新的体外节律诱导剂以研究C6神经胶质瘤细胞近日节律基因的表达情况。方法采用PMA（phorbol 12-myristate 13-acetate），100nmol／L）及50％的马血清刺激体外培养的小鼠NIH-3T3细胞。RT-PCR检测不同时间点mPer1 mRNA的表达水平，比较两者的诱导效率；PMA刺激体外培养的大鼠C6神经胶质瘤细胞，RT—PCR检测不同时间点rPer1，rDbp mRNA的表达水平。结果PMA及50％的马血清均能诱导NIH-3T3细胞mPer1的近日节律表达且诱导效率近似；并首次发现C6细胞的rPer1，rDbp基因存在着近日节律振荡。结论PMA是一种有效的体外节律诱导剂；经PMA诱导后，C6细胞的节律基因存在近日振荡，为体外研究授时因子对近日节律的导引机制提供依据。

PMA-induced Circadian Gene Expression of rPer1,rDbp in C6 Cells

Objective To investigate the expression level of circadian gene in C6 glioma cells by searching for a novel rhythm-inductor in vitro. Method In order to compare the induction efficiency of PMA with horse serum,the cultured NIH-3T3 cells were stimulated by PMA(phorbol 12-myristate 13-acetate ,100 nmol/L) and 50% horse serum respectively and performed RT-PCR to examine the expression of mPer1 mRNAs at the indicated times. The cultured C6 glioma cells were also stimulated by PMA and examined the expression of rPer1, rDbp mRNAs at the indicated times by RT-PCR. Result Both PMA and 50% horse serum could induce the circadian gene expression of mPer1 in NIH-3T3 cells and achieved similar induction efficiency ,and it was confirmed that circadian oscillation of rPer1,rDbp existed in C6 glioma cells. Conclusion PMA is an effective rhythm inductor in vitro, and elicit the circadian oscillation of clock genes in C6 glioma cells, which confer an advantage to the investigation of circadian entrainment mechanisms in vitro.