EB病毒感染对IGF2基因甲基化和印迹表达影响的研究

发布时间：2018-05-27 19:59

本文选题：胰岛素样生长因子2 + EB病毒　；参考：《青岛大学》2016年硕士论文

【摘要】：目的:明确EBV感染对胰岛素样生长因子2(insulin-like growth factor2,IGF2)编码基因启动子区甲基化、印迹状态及表达的影响,探讨IGF2基因与EBVa GC发生发展的关系。方法:亚硫酸氢盐基因组测序法(Bisufite genomic sequencing,BGS)检测EBV阳性和阴性细胞系以及EBV相关胃癌(EBV-associated gastric carcinoma,EBVa GC)和EBV阴性胃癌(EBV-negatived gastric carcinoma,EBVn GC)组织中IGF2基因启动子区甲基化状态。聚合酶链式反应-限制性片段长度多态性(polymerase chain reaction-restriction fragment length polymorphism,PCR-RFLP)分析检测EBVa GC和EBVn GC及癌旁组织标本中IGF2基因印迹状态。实时荧光定量PCR(Real time quantitative PCR,real time q PCR)检测EBVa GC和EBVn GC组织中IGF2 m RNA的转录表达;免疫组化技术检测EBVa GC,EBVn GC及癌旁组织中IGF2蛋白表达。结果:(1)BGS结果显示EBV阳性和阴性细胞系中IGF2基因均为高甲基化;EBVa GC组织平均甲基化率为38.17%,EBVn GC组织平均甲基化率为23.06%,统计分析显示EBVa GC组织中IGF2基因启动子甲基化水平明显高于EBVn GC组织(P0.05)。(2)EBVa GC组织中IGF2印迹缺失率为61.3%(61.3%,19/31);EBVn GC组织中IGF2印迹缺失率为33.3%(33.3%,15/45)。EBVa GC组织中IGF2印迹缺失率高于EBVn GC组织,两者存在显著性差异(χ2=5.803,P0.05);(3)EBVa GC和EBVn GC组织中IGF2 m RNA转录表达无显著性差异(P0.05)。(4)IGF2蛋白表达在EBVa GC和EBVn GC组织中无显著差异(P0.05);胃癌组织中IGF2蛋白明显表达高于癌旁组织(P0.05)。结论(1)EBV感染可诱发IGF2基因启动子的高甲基化和IGF2基因印迹丢失。(2)未观察到EBV感染、IGF2基因甲基化和印迹状态对IGF2转录和表达的影响,表明IGF2表达的调控机制较为复杂,可能有多种因素的共同调节。(3)胃癌组织中IGF2蛋白的高表达表明其可能参与胃癌的发生发展。
[Abstract]:Aim: to investigate the effect of EBV infection on methylation, imprinting status and expression of insulin-like growth factor 2(insulin-like growth factor2IGF2 (insulin-like growth factor 2) gene promoter, and to explore the relationship between IGF2 gene and EBVa GC. Methods: Bisufite genomic sequencing method was used to detect the methylation status of IGF2 promoter region in EBV associated gastric cancer cell lines, EBV-associated gastric carcinoma-associated gastric (EBVa GCC) and EBV negative gastric carcinomatous gastric (EBVn GCCs). Polymerase chain reaction-restriction fragment length polymorphisms were analyzed by polymerase chain reaction-restriction fragment length polymorphism (RFLP) to detect the IGF2 gene imprinting status in EBVa GC and EBVn GC and adjacent tissues. The expression of IGF2 m RNA in EBVa GC and EBVn GC was detected by real-time fluorescence quantitative PCR(Real time quantitative real time q PCR), and the expression of IGF2 protein in EBVa GC and adjacent tissues was detected by immunohistochemistry. Results the results of EBV positive and negative cell lines showed that the average methylation rate of IGF2 gene was 38.17%. The average methylation rate of IGF2 gene promoter in EBVa GC tissue was 23.06. Statistical analysis showed that IGF2 gene promoter was methylated water in EBVa GC tissue. The deletion rate of IGF2 blotting in GC tissues was significantly higher than that in EBVn GC tissues (P 0.05U. 2). The deletion rate of IGF2 blotting in GC tissues was significantly higher than that in GC tissues. The deletion rate of IGF2 blotting in GC tissues was 33.3% higher than that in GC tissues of EBVn GC. The deletion rate of IGF2 blotting in GC tissues was significantly higher than that in GC tissues of EBVn GC, and the deletion rate of IGF2 blotting in GC tissues was higher than that in GC tissues of EBVn GC. There was no significant difference in the expression of IGF2 m RNA between GC and EBVn. There was no significant difference in the expression of IGF2 m RNA between EBVa GC and EBVn GC, and the expression of IGF2 protein in gastric cancer was significantly higher than that in adjacent tissues. Conclusion the hypermethylation of IGF2 promoter and the loss of IGF2 gene imprinting were induced by IGF2 infection. The effect of EBV infection on IGF2 transcription and expression was not observed, indicating that the regulation mechanism of IGF2 expression was complicated. The overexpression of IGF2 protein in gastric cancer tissues suggests that it may be involved in the development of gastric cancer.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R735.2

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