文献类型：期刊文献

中文题名：HPV E7在人食管鳞癌细胞系EC9706中基因沉默的研究

英文题名：-A study on RNA silencing of HPV E7 gene in human esophageal squamous cancer cell line EC9706

作者：武双[1];刁明坤[3];李劲涛[2];刘洪伟[3];李凡[2];龚平[1];钟儒刚[2];曾毅[3]

第一作者：武双

机构：[1]北京联合大学生物化学工程学院,100023;[2]北京工业大学生命科学与生物工程学院环境与病毒肿瘤学北京市重点实验室;[3]中国疾病预防控制中心病毒病预防控制所 传染病预防控制国家重点实验室

第一机构：北京联合大学生物化学工程学院

年份：2015

卷号：29

期号：2

起止页码：130-132

中文期刊名：中华实验和临床病毒学杂志

外文期刊名：Chinese Journal of Experimental and Clinical Virology

收录：CSTPCD;;Scopus;CSCD:【CSCD2015_2016】；PubMed;

基金：传染病预防控制国家重点实验室自主研究重点课题（2011SKLID103）;国家科技支撑计划项目（No.2006BA119803）;国家863计划课题（2012AA02A404）;北京联合大学新起点计划项目

语种：中文

中文关键词：乳头状瘤病毒;人;基因;食管肿瘤;细胞系

外文关键词：Papillomavirus,human;Genes;Esophageal neoplasms;Call line

摘要：目的 探讨基因沉默人乳头瘤病毒（human papillomavirus,HPV）阳性食管鳞癌细胞系EC9706中E7基因后对细胞内基因表达的影响.方法 使用RNAi专用逆转录病毒载体构建可干扰HPV18 E7基因的质粒,转入EC9706细胞系,筛选获得稳定表达干扰片段的细胞株.使用Real-timePCR和Western Blot对胞内相关基因和蛋白进行检测,并比较细胞活力差异.结果 通过质粒构建与转染,获得了两株稳定表达干扰片段的细胞株EC9706R8-1及EC9706R8-4.Real-time PCR检测结果发现病毒基因E6、E7均被下调.Western Blot检测结果表明在病毒蛋白E7通路中控制细胞周期的p27蛋白表达量有显著的提高.同时,E6及E7基因下调后,细胞活力也明显下降.结论 EC9706食管癌细胞系中HPV18 E7基因的沉默很可能与食管上皮细胞的癌变有关,该结果为HPV相关食管癌的基因沉默治疗研究提供了一定的理论基础.
Objective To explore on the impact of the integrated HPV type 18 on carcinogenesis of esophageal squamous cancer cell line EC9706.Methods A E7 gene silencing sequence was induced into the pSUPER.retro.neo ＋ gfp vector to down regulate the viral mRNA in EC9706 cell lines.Real time PCR and Western Blot were employed to detect the related genes in both mRNA and protein level respectively in these cell lines.The cell viability was also detected by the proliferate property.Results The E6 and E7 gene down regulated of stable cell lines were obtained.The Western Blot results also showed that there was a huge increase in the p27 protein which involved in the E7-cellular protein network.The reduced cell viability showed that the expressed viral genes in EC9706 cell lines are the factors to enhance the cell viability.Conclusion This study proved the integrated HPV could be the pathogenetic factor of carcinogenesis in esophageal squamous cancer cell line EC9706,and the promising results established the basis of RNA silencing in therapeutic practice in HPV induced cancer.