本文選題：胰島素樣生長因子2 + EB病毒　； 參考：《青島大學(xué)》2016年碩士論文

【摘要】：目的:明確EBV感染對胰島素樣生長因子2(insulin-like growth factor2,IGF2)編碼基因啟動子區(qū)甲基化、印跡狀態(tài)及表達的影響,探討IGF2基因與EBVa GC發(fā)生發(fā)展的關(guān)系。方法:亞硫酸氫鹽基因組測序法(Bisufite genomic sequencing,BGS)檢測EBV陽性和陰性細胞系以及EBV相關(guān)胃癌(EBV-associated gastric carcinoma,EBVa GC)和EBV陰性胃癌(EBV-negatived gastric carcinoma,EBVn GC)組織中IGF2基因啟動子區(qū)甲基化狀態(tài)。聚合酶鏈式反應(yīng)-限制性片段長度多態(tài)性(polymerase chain reaction-restriction fragment length polymorphism,PCR-RFLP)分析檢測EBVa GC和EBVn GC及癌旁組織標本中IGF2基因印跡狀態(tài)。實時熒光定量PCR(Real time quantitative PCR,real time q PCR)檢測EBVa GC和EBVn GC組織中IGF2 m RNA的轉(zhuǎn)錄表達;免疫組化技術(shù)檢測EBVa GC,EBVn GC及癌旁組織中IGF2蛋白表達。結(jié)果:(1)BGS結(jié)果顯示EBV陽性和陰性細胞系中IGF2基因均為高甲基化;EBVa GC組織平均甲基化率為38.17%,EBVn GC組織平均甲基化率為23.06%,統(tǒng)計分析顯示EBVa GC組織中IGF2基因啟動子甲基化水平明顯高于EBVn GC組織(P0.05)。(2)EBVa GC組織中IGF2印跡缺失率為61.3%(61.3%,19/31);EBVn GC組織中IGF2印跡缺失率為33.3%(33.3%,15/45)。EBVa GC組織中IGF2印跡缺失率高于EBVn GC組織,兩者存在顯著性差異(χ2=5.803,P0.05);(3)EBVa GC和EBVn GC組織中IGF2 m RNA轉(zhuǎn)錄表達無顯著性差異(P0.05)。(4)IGF2蛋白表達在EBVa GC和EBVn GC組織中無顯著差異(P0.05);胃癌組織中IGF2蛋白明顯表達高于癌旁組織(P0.05)。結(jié)論(1)EBV感染可誘發(fā)IGF2基因啟動子的高甲基化和IGF2基因印跡丟失。(2)未觀察到EBV感染、IGF2基因甲基化和印跡狀態(tài)對IGF2轉(zhuǎn)錄和表達的影響,表明IGF2表達的調(diào)控機制較為復(fù)雜,可能有多種因素的共同調(diào)節(jié)。(3)胃癌組織中IGF2蛋白的高表達表明其可能參與胃癌的發(fā)生發(fā)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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R735.2

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