转录因子SP1调控小鼠前成骨细胞中MEPE基因表达的研究

The Study of Transcription Factor SP1 Regulating MEPE Gene Expression in Preosteoblasts of Mouse

克隆转录因子SP1并构建其真核表达载体;在双荧光素酶基因检测报告系统下观察3种MAPK信号通路抑制剂调控SP1对不同长度的MEPE基因表达的调控作用,以分析转录因子SP1作用于MEPE基因的显著区段,并利用实时定量RT-PCR法分析SP1调控MEPE基因的表达。根据双荧光素酶基因检测报告系统的研究表明在小鼠前成骨细胞中,SP1可以抑制MEPE基因的转录活性,在(-1 081~66)1 147 bp片段区域内抑制作用较为显著,SP1可通过抑制JNK途径抑制MEPE基因的转录活性;实时定量RT-PCR法分析再次验证SP1抑制MEPE基因的表达水平。这表明转录因子SP1对MEPE基因表达具有显著的调节作用,从而表明转录因子SP1在骨形成发育过程中起到重要的生物学意义。

Transcription factor SP1 was cloned and the recombinant plasmid of eukaryotic expression vector of SP1 was also constructed. Then the dual luciferase reporter assay was used to observe the effects of SP1 in the MEPE genes of different lengths by 3 MAPK signaling pathway inhibitors in order to determine the significant effect region of SP1 on MEPE. Moreover the real time PCR was used to analyze the expression activity of MEPE gene regulated by SP1. Dual luciferase reporter assay showed that SP1 could inhibit the transcription activity of MEPE gene in preosteoblasts of mouse, especially in the fragment area of （-1 081-66） 1 147 bp. SP1 could inhibit the transcription activity of MEPE gene by controlling the JNK MAPK single pathway. The Real time PCR verified again that SP1 prevented the expression activity of MEPE gene. Therefore we could conclude that SP1 had remarkable regulation effect on the expression of the MEPE gene, so as it had the important biological significance in the process of bone development.