ezrin基因在几种癌细胞中的表达及其5′侧翼区序列转录活性的鉴定

doi:10.3971/j.issn.1000-8578.1222

ezrin基因在几种癌细胞中的表达及其5′侧翼区序列转录活性的鉴定

Expression of ezrin Gene in Some Carcinoma Cells and Identification of Its 5′-Flanking Transcriptional Activity

摘要

摘要: 目的检测ezrin基因在几种癌细胞中的表达及其5′侧翼区序列的转录活力,探讨ezrin基因在癌细胞的表达调控机制。方法采用逆转录聚合酶链反应(RT-PCR)及Western blot法检测ezrin基因在食管癌细胞EC109、EC171、EC8712、SHEEC,胃癌细胞N87、BGC823,肺癌细胞A549、95D,肝癌细胞HepG2,白血病细胞K562和宫颈癌细胞HeLa中的mRNA和蛋白表达水平;采用PCR法构建以ezrin基因翻译起始位点上游-1444/+134序列为启动子的真核细胞表达质粒pGLB-hE(-1444/+134);采用双荧光素酶报告基因分析系统检测-1444/+134序列在EC109、Hela、A549和BGC823细胞中的转录活力。结果ezrin基因在食管癌等几种癌细胞中均有高表达,其5′侧翼区-1444/+134序列具有较强的转录活力。结论ezrin基因5′侧翼区序列的转录活性可能对ezrin基因的几种癌细胞中的高表达起重要作用。

Abstract: Objective To investigate the expression of ezrin gene in some carcinoma cells,to detect the transcriptional activity of its 5′-flanking region,and to elucidate the transcriptional regulatory mechanism of the ezrin gene in carcinoma cells. Methods Expression of ezrin gene in esophageal carcinoma EC109,EC171,EC8712 and SHEEC,gastric carcinoma N87 and BGC823,lung carcinoma A549 and 95D,hepatocellular carcinoma HepG2,leukemia K562,and cervical carcinoma HeLa cells was detected using reverse transcriptase polymerase chain reaction (RT2PCR) and western blot analysis. Plasmid p GLB2hE( -1444/ + 134), containing - 1444/ + 134 of ez rin gene fused to the luciferase gene, was generated by PCR method. Transcriptional activity of the - 1444/ + 134 region in some carcinoma cells was examined using dual2luciferase reporter assay system. Results 　All the tested carcinoma cells high2expressed ez rin mR2 NA and Ezrin protein. - 1444/ + 134 region of ez rin gene had st rong t ranscriptional activity compared with SV40 promoter ( P < 0. 05) in EC109, Hela, A549 and BGC823 cells. Conclusion 　The t ranscrip2 tional activity of the ez rin gene

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