EBV相关胃癌及鼻咽癌中病毒潜伏期基因启动子甲基化状态的研究
发布时间：2018-10-31 21:07
【摘要】： EB病毒(Epstein-Barr virus, EBV)是重要的DNA肿瘤病毒,属人类疱疹病毒科γ疱疹病毒亚科成员,与人类多种恶性肿瘤密切相关。EBV相关胃癌(EBV-associated gastric carcinoma, EBVaGC)及鼻咽癌组织中病毒基因表现为限制性表达,可能与病毒利用表观遗传机制,尤其是利用DNA甲基化来调控病毒基因启动子的活性。研究表明,EBV相关肿瘤组织中病毒可借甲基化沉默部分基因,导致相关蛋白表达水平的降低,因而对宿主免疫系统的刺激也减弱,病毒借此来逃避宿主免疫攻击,同时也提示针对病毒基因表观改变的靶向去甲基化作用有可能成为治疗EBV阳性肿瘤的新方法。 目的研究分析青岛地区EBVaGC及鼻咽癌组织中EBV主要潜伏期编码基因启动子甲基化状态。 方法选择32例EBVaGC及20例EBV阳性鼻咽癌(nasopharyngeal carcinoma,NPC)作为研究对象,以EBV阳性细胞系Raji, B95-8和Akata作为对照,采用甲基化特异性PCR (Methylation-Specific PCR, MSP)及亚硫酸氢盐-基因组测序法(Bisulfite genomic sequencing, BGS)检测分析其EBV核抗原编码基因启动子Cp,Wp和Qp以及EBV潜伏膜蛋白LMP1和LMP2A编码基因启动子的甲基化状态。 结果①EBV阳性细胞系中EBV编码基因启动子甲基化状态表现为：B95-8细胞系Cp未发生甲基化,Raji细胞系Cp呈高甲基化,但在Akata细胞系同时检测到甲基化和未甲基化的Cp。Raji和Akata细胞系Wp呈高甲基化,但在B95-8细胞系中则表现为部分未甲基。3种细胞系中均未检测到Qp、LMPlp和LMP2Ap的甲基化。②EBVaGC及鼻咽癌肿瘤标本中EBV核抗原编码基因启动子的甲基化状态与肿瘤细胞的潜伏感染类型基本一致：Cp和Wp呈高甲基化状态,而Qp未发生甲基化。③EBVaGC和EBV阳性鼻咽癌组织中LMP1和LMP2A编码基因启动子甲基化状态有差别,二者在EBVaGC组织中的甲基化程度明显高于EBV阳性鼻咽癌(LMP1：χ2=19.1462, P0.0001; LMP2Ap:χ2=11.0139, P=0.0009)。 结论EBV相关胃癌和EBV阳性鼻咽癌组织中病毒潜伏期基因启动子的甲基化状态不同,甲基化是调控EBV潜伏期基因启动子活动的重要机制之一。
[Abstract]:EB virus (Epstein-Barr virus, EBV) is an important DNA tumor virus, belonging to the subfamily of human herpesviridae 纬 herpesvirus. EBV associated gastric cancer (EBV-associated gastric carcinoma,) is closely related to various human malignant tumors. EBVaGC) and nasopharyngeal carcinoma (NPC) tissues, which may be associated with the use of epigenetic mechanisms, especially the use of DNA methylation to regulate the activity of viral gene promoters. Studies have shown that viruses in tumor tissues associated with EBV can silence some genes by methylation, leading to a decrease in the level of expression of related proteins, which weakens the stimulation of the host immune system, thereby evading the host immune attack. It is also suggested that the targeted demethylation targeting the epigenetic changes of virus genes may be a new method for the treatment of EBV positive tumors. Objective to study the methylation status of promoter of EBV major latent gene in EBVaGC and nasopharyngeal carcinoma tissues in Qingdao. Methods 32 cases of EBVaGC and 20 cases of EBV positive nasopharyngeal carcinoma (nasopharyngeal carcinoma,NPC) were studied. The EBV positive cell lines Raji, B95-8 and Akata were used as control, and the methylation specific PCR (Methylation-Specific PCR,) was used. MSP) and (Bisulfite genomic sequencing, BGS) were used to detect the methylation status of the promoter of EBV nuclear antigen encoding gene Cp,Wp and Qp, as well as EBV latent membrane protein LMP1 and LMP2A gene promoter. Results the methylation status of the promoter of EBV coding gene in 1EBV positive cell lines was as follows: no methylation occurred in B95-8 cell line Cp, but hypermethylation in Raji cell line Cp. However, hypermethylation of both methylated and unmethylated Cp.Raji and Akata Wp was detected in Akata cell line, but Qp, was not detected in B95-8 cell line. Methylation of LMPlp and LMP2Ap. The methylation status of EBV nucleoantigen gene promoter in 2EBVaGC and nasopharyngeal carcinoma samples was basically consistent with the latent infection type of tumor cells: Cp and Wp were hypermethylated. However, there was no methylation in Qp. The methylation status of LMP1 and LMP2A gene promoter in 3EBVaGC and EBV positive nasopharyngeal carcinoma tissues was significantly higher than that in EBV positive nasopharyngeal carcinoma tissues (LMP1: 蠂 2, 19.1462, P 0.0001). LMP2Ap: 蠂 2: 11.0139, P = 0.0009). Conclusion the methylation status of viral latent gene promoter in EBV associated gastric cancer and EBV positive nasopharyngeal carcinoma is different. Methylation is one of the important mechanisms that regulate the promoter activity of EBV latency gene.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R735.2;R739.63
本文编号：2303553
[Abstract]:EB virus (Epstein-Barr virus, EBV) is an important DNA tumor virus, belonging to the subfamily of human herpesviridae 纬 herpesvirus. EBV associated gastric cancer (EBV-associated gastric carcinoma,) is closely related to various human malignant tumors. EBVaGC) and nasopharyngeal carcinoma (NPC) tissues, which may be associated with the use of epigenetic mechanisms, especially the use of DNA methylation to regulate the activity of viral gene promoters. Studies have shown that viruses in tumor tissues associated with EBV can silence some genes by methylation, leading to a decrease in the level of expression of related proteins, which weakens the stimulation of the host immune system, thereby evading the host immune attack. It is also suggested that the targeted demethylation targeting the epigenetic changes of virus genes may be a new method for the treatment of EBV positive tumors. Objective to study the methylation status of promoter of EBV major latent gene in EBVaGC and nasopharyngeal carcinoma tissues in Qingdao. Methods 32 cases of EBVaGC and 20 cases of EBV positive nasopharyngeal carcinoma (nasopharyngeal carcinoma,NPC) were studied. The EBV positive cell lines Raji, B95-8 and Akata were used as control, and the methylation specific PCR (Methylation-Specific PCR,) was used. MSP) and (Bisulfite genomic sequencing, BGS) were used to detect the methylation status of the promoter of EBV nuclear antigen encoding gene Cp,Wp and Qp, as well as EBV latent membrane protein LMP1 and LMP2A gene promoter. Results the methylation status of the promoter of EBV coding gene in 1EBV positive cell lines was as follows: no methylation occurred in B95-8 cell line Cp, but hypermethylation in Raji cell line Cp. However, hypermethylation of both methylated and unmethylated Cp.Raji and Akata Wp was detected in Akata cell line, but Qp, was not detected in B95-8 cell line. Methylation of LMPlp and LMP2Ap. The methylation status of EBV nucleoantigen gene promoter in 2EBVaGC and nasopharyngeal carcinoma samples was basically consistent with the latent infection type of tumor cells: Cp and Wp were hypermethylated. However, there was no methylation in Qp. The methylation status of LMP1 and LMP2A gene promoter in 3EBVaGC and EBV positive nasopharyngeal carcinoma tissues was significantly higher than that in EBV positive nasopharyngeal carcinoma tissues (LMP1: 蠂 2, 19.1462, P 0.0001). LMP2Ap: 蠂 2: 11.0139, P = 0.0009). Conclusion the methylation status of viral latent gene promoter in EBV associated gastric cancer and EBV positive nasopharyngeal carcinoma is different. Methylation is one of the important mechanisms that regulate the promoter activity of EBV latency gene.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R735.2;R739.63
【参考文献】
相关期刊论文 前4条
1 王云,罗兵,赵鹏,黄葆华;EB病毒相关胃癌组织中病毒基因的表达[J];癌症;2004年07期
2 刘振声，李宝民，刘彦仿，，曾毅;EB病毒与促癌物协同作用诱发人鼻咽恶性淋巴瘤和未分化癌的研究[J];病毒学报;1996年01期
3 王琦;胡凤成;李春明;;DNA甲基化研究进展[J];科技情报开发与经济;2009年29期
4 周晓彬;纪新强;徐莉;;医用统计学软件PPMS 1.5的组成和应用特点[J];齐鲁医学杂志;2009年01期
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