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目的:研究鼻咽癌细胞中ezrin基因启动子上游序列的转录调控特性。方法:构建一系列携带ezrin基因-1541/-706序列的报告基因表达载体,ezrin基因-1541/-706序列以正向和反向分别连接至不含启动子的报告基因上游、ezrin启动子上游、SV40启动子上游、ezrin启动子或SV40启动子控制的报告基因下游,将质粒转染CNE2细胞,检测荧光素酶活性。结果:CNE2细胞中,当-1541/-706序列正向位于报告基因上游时表现出启动子活性,其转录激活作用约为ezrin启动子的50%;反向连接时无启动子活性。而且,当-1541/-706序列正向位于ezrin启动子或SV40启动子上游时,显著提高荧光素酶表达;当反向位于启动子下游、正向或反向位于启动子控制的报告基因下游时,转录增强作用消失。结论:CNE2细胞中ezrin基因启动子上游序列具有转录激活和转录增强作用,这种作用具有DNA序列位置和方向依赖性。
Objective: To study the transcriptional regulation of ezrin gene upstream of nasopharyngeal carcinoma cells. METHODS: A series of reporter gene expression vectors carrying the ezrin gene -1541 / -706 sequence were constructed. The ezrin gene -1541 / -706 sequence was ligated in the forward and reverse directions respectively to the upstream of the promoter-free reporter and upstream of the ezrin promoter , Upstream of SV40 promoter, downstream of ezrin promoter or SV40 promoter, the plasmid was transfected into CNE2 cells to detect luciferase activity. RESULTS: In CNE2 cells, the promoter sequence of -1541 / -706 was located upstream of the reporter gene and its transcriptional activation was about 50% of that of the ezrin promoter. No promoter activity was detected in the reverse connection. Moreover, luciferase expression was significantly increased when the -1541 / -706 sequence was located directly upstream of the ezrin promoter or the SV40 promoter; when the reverse orientation was downstream of the promoter and the forward or reverse orientation was downstream of the promoter-controlled reporter When the transcription enhancement disappears. CONCLUSION: The upstream of ezrin gene promoter in CNE2 cells has transcriptional activation and transcriptional enhancement, which has the DNA sequence location and direction-dependent.