本文選題：胰島素樣生長(zhǎng)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(duì)胰島素樣生長(zhǎng)因子2(insulin-like growth factor2,IGF2)編碼基因啟動(dòng)子區(qū)甲基化、印跡狀態(tài)及表達(dá)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(cè)序法(Bisufite genomic sequencing,BGS)檢測(cè)EBV陽(yáng)性和陰性細(xì)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(dòng)子區(qū)甲基化狀態(tài)。聚合酶鏈?zhǔn)椒磻?yīng)-限制性片段長(zhǎng)度多態(tài)性(polymerase chain reaction-restriction fragment length polymorphism,PCR-RFLP)分析檢測(cè)EBVa GC和EBVn GC及癌旁組織標(biāo)本中IGF2基因印跡狀態(tài)。實(shí)時(shí)熒光定量PCR(Real time quantitative PCR,real time q PCR)檢測(cè)EBVa GC和EBVn GC組織中IGF2 m RNA的轉(zhuǎn)錄表達(dá);免疫組化技術(shù)檢測(cè)EBVa GC,EBVn GC及癌旁組織中IGF2蛋白表達(dá)。結(jié)果:(1)BGS結(jié)果顯示EBV陽(yáng)性和陰性細(xì)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(jì)分析顯示EBVa GC組織中IGF2基因啟動(dòng)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(dá)無(wú)顯著性差異(P0.05)。(4)IGF2蛋白表達(dá)在EBVa GC和EBVn GC組織中無(wú)顯著差異(P0.05);胃癌組織中IGF2蛋白明顯表達(dá)高于癌旁組織(P0.05)。結(jié)論(1)EBV感染可誘發(fā)IGF2基因啟動(dòng)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(duì)IGF2轉(zhuǎn)錄和表達(dá)的影響,表明IGF2表達(dá)的調(diào)控機(jī)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(dá)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(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：R735.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前4條

1 陳文芳;徐惠霞;紀(jì)恒勝;李玉軍;;IGF2和IMP3在胃癌組織中的表達(dá)及其意義[J];齊魯醫(yī)學(xué)雜志;2015年04期

2 黃健;孫朝暉;鄒軍榮;朱曉艷;熊英俊;;HBV轉(zhuǎn)染肝癌細(xì)胞系基因表達(dá)譜改變的研究[J];重慶醫(yī)學(xué);2011年11期

3 Philippe Couvert;Alain Carrié;Jacques Pariès;Jenny Vaysse;Audrey Miroglio;Antoine Kerjean;Pierre Nahon;Jamel Chelly;Jean-Claude Trinchet;Michel Beaugrand;Nathalie Ganne-Carrié;;Liver insulin-like growth factor 2 methylation in hepatitis C virus cirrhosis and further occurrence of hepatocellular carcinoma[J];World Journal of Gastroenterology;2008年35期

4 劉重陽(yáng);陳東風(fēng);王軍;;乙型肝炎病毒X基因-丙型肝炎病毒C基因共表達(dá)對(duì)肝癌細(xì)胞胰島素樣生長(zhǎng)因子-2表達(dá)的影響[J];胃腸病學(xué)和肝病學(xué)雜志;2008年05期

，

本文編號(hào)：1943554