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Transcriptional Regulatory Properties of Human ezrin Gene Enhancer in Several Carcinoma Cells
Zhang Qingfeng1, Wei Jinqi2, Zhang Fangting3, Shao Min1, Shen Huifang1, Gao Shuying1,3*
1Department of Biochemistry and Moleculuar Biology, Zhuhai Campus of Zunyi Medical University, Zhuhai 519041, China; 2Department of Gastroenterology, the Fifth Affiliated Hospital of Sun Yat-sen University, Zhuhai 519000, China;
Abstract: To investigate transcriptional regulatory properties of ezrin gene enhancer in carcinoma cells， Ezrin expression in human esophageal carcinoma EC109， nasopharyngeal carcinoma CNE2 and cervical carcinoma HeLa cells was detected by Western blot. A series of reporter gene expression vectors carrying ezrin enhancer–1541/–706 sequence were constructed using DNA fragments orientating clone method and then transfected into EC109， CNE2 and HeLa cells for luciferase assay. It was found that the expression levels of Ezrin in the detected three cell lines were not obviously different. In EC109 cells， when the ezrin –1541/–706 segment was located upstream of luc reporter gene without promoter in the forward orientation， it exhibited transcriptional activation like a promoter; while this transactivation was nearly abolished when this segment was reversed. When this segment was located upstream of the ezrin promoter or SV40 promoter in the forward orientation， it dramatically increased luciferase expression. However， the transcriptional enhancement disappeared when this segment was located upstream of promoters in the reverse orientation， or downstream of reporter genes in the forward or reverse orientation. The transcriptional regulation of ezrin –1541/–706 segment in CNE2 and HeLa cells was partly similar， but not completely identical to that in EC109 cells. These data suggested that the ezrin enhancer could exhibit transcriptional activation and enhancement， in a position/orientation-dependent and cell-type-specific manner.
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