Site-directed analysis of key transcriptional regulatory regions of ezrin gene in HeLa cells / 肿瘤

ABSTRACT

Objective: To identify key transcriptional regulatory regions of ezrin gene in HeLa cells, thus this work maked a preliminary study for revealing its transcriptional regulatory mechanism. Methods: The recombinant pGLB plasmids containing different lengths of DNA fragments upstream of translation initiation site of ezrin gene as reporter promoters were constructed using nested-deletion method, and the promoter activities were detected using dual-luciferase reporter assay system. The potential transcription factor binding sites at key transcriptional regulatory region of ezrin gene were predicted using on-line analyzing programme. Results: The recombinant pGLB plasmids containing different lengths of 5′-flanking region of ezrin gene were obtained. When the lengths of ezrin 5′-flanking region were reduced from - 1 324 to - 890, the transcriptional activity decreased by about 80%. If the length of 5′-flanking region were deleted from - 146 to - 32, the transcriptional activities were nearly abolished. Sp 1 transcriptional factor binding sites were ubiquitous at the - 1 324/ - 890 and - 146/ - 32 regions. Conclusion: The - 1 324/ - 890 and - 146/ - 32 regions were two key transcriptional regulatory regions of ezrin gene in HeLa cells. Sp 1 may be an important factor for regulating the transcription of ezrin gene.