miR-127、miR-155和miR-200a在EB病毒感染相关的儿童散发性伯基特淋巴瘤中的表达及意义

发布时间：2018-06-01 23:24

本文选题：伯基特淋巴瘤 + EB病毒　；参考：《南昌大学》2014年硕士论文

【摘要】：目的：探讨miR-127、miR-155和miR-200a在EB病毒（Epstein-Barr virus,EBV）感染相关的儿童散发性伯基特淋巴瘤(Burkitt's lymphoma,BL)中的表达及意义。方法： 1、收集2005～2012年江西省儿童医院病理科外科手术切除的40例BL标本及15例慢性扁桃体炎标本。分析其临床病理特征及随访资料，制备组织芯片，运用原位杂交技术对标本进行EBV基因检测；根据EBV基因检测结果将BL分为EBV感染阴性组、EBV感染阳性组和正常对照组。 2、运用qRT-PCR法，对已分组的40例BL和15例慢性扁桃体炎石蜡标本进行miR-127、 miR-155和miR-200a表达的检测。 3、培养BL细胞株Raji（EBV阳性）和Ramos（EBV阴性），运用qRT-PCR法对细胞株中miR-127、 miR-155和miR-200a的表达进行检测。 4、运用荧光原位杂交（fluorescence in-situ hybridization,FISH）技术检测40例BL石蜡组织及BL细胞株中c-myc基因的异常情况。 5、使用13.0统计软件包对实验数据进行分析,计量资料采用“平均值±标准差”,计数资料采用t检验。P0.05为差异有统计学意义。结果： 1、40例BL病例中16例EBER阳性（40%），其中2例（12.5%）位于肠系膜淋巴结、4例（25%）位于回盲部肠组织、10例（62.5%）位于肠系膜+回盲部；24例EBER阴性（60%），其中2例(8.33%)位于肠系膜淋巴结、6例(25%)位于回盲部肠组织、16例(66.67%)位于肠系膜+回盲部。15例扁桃体组织EBER均为阴性（100%） 2、40例BL中38例c-myc基因断裂阳性（95%），其中EBER阴性23例（60.53％），EBER阳性15例（39.47%）；2例c-myc基因信号正常（5％），EBER阳性和EBER阴性各1例（各50%）。Raji细胞株c-myc基因断裂阳性，Ramos细胞株基因信号正常。 3、与EBER阴性的BL病例及扁桃体组织比较，EBER阳性的BL病例miR-127表达上调（P㩳0.05），差异有统计学意义。Raji细胞株和Ramos细胞株相比，Raji细胞株中miR-127表达上调（P㩳0.05）。与EBER阴性的BL病例比较，，miR-155在EBER阳性的BL病例中表达上调，但无显著差异性（P㧐0.05），EBER阳性BL的病例与扁桃体组织相比表达上调（P㩳0.05），差异有统计学意义。Raji和Ramos比较，在Raji细胞株中miR-155的表达上调（P㩳0.05）。miR-200a的表达在EBV阳性的BL相比EBV阴性的BL(0.504±1.214)和扁桃体组织表达显著降低（P㩳0.05），Raji和Ramos比较miR-200a表达显著降低（P㩳0.05）。miR-127、miR-155和miR-200a的表达与c-myc基因比较均无统计学意义（P㧐0.05）结论：（1）miR-127、miR-155和miR-200a在EBV不同感染状态的儿童散发性BL中表达存在差异。（2）miR-127、miR-155的高表达及miR-200a的低表达与儿童散发性BL的发病机制及EBV感染密切相关，可能起着类似癌基因/抑癌基因的作用。
[Abstract]:Objective: To investigate the expression and significance of miR-127 miR-155 and miR-200a in children with Epstein-Barr virus (EBV) infection. Methods: 1. From 2005 to 2012, 40 cases of BL and 15 cases of chronic tonsillitis were collected. The clinicopathological characteristics and follow-up data were analyzed, tissue microarray was prepared, and EBV gene was detected by in situ hybridization. BL was divided into two groups according to the results of EBV gene detection: positive EBV infection group and normal control group. 2. The expression of miR-127, miR-155 and miR-200a in 40 cases of BL and 15 cases of chronic tonsillitis were detected by qRT-PCR method. The expression of miR-127, miR-155 and miR-200a was detected by qRT-PCR method. (4) fluorescence in situ hybridization (fish) technique was used to detect the abnormality of c-myc gene in 40 cases of BL paraffin tissues and BL cell lines. 5. The experimental data were analyzed with 13.0 statistical software package. The data were measured by "mean 卤standard deviation" and counted by t test. P05 was statistically significant. Results: 1of the 40 cases of BL, 16 cases were positive for EBER, of which 2 cases were located in mesenteric lymph nodes, 4 cases were located in mesenteric lymph nodes, 10 cases were located in ileocecal intestinal tissue, 10 cases were located in ileocecal ileocecum, 24 cases were located in ileocecal ileocecum, and 2 cases were located in 6 cases of mesenteric lymph nodes, among them, 2 cases were located in mesenteric lymph nodes, 6 cases were located in mesenteric lymph nodes, and 2 cases were located in mesenteric lymph nodes. In ileocecal ileocecum, 16 cases were located in ileocecal jejunum (66.67). In mesenteric ileocecal part, 15 cases of tonsil tissue EBER were all negative. 2among 40 cases of BL, 38 cases were positive for c-myc gene breakage. Among them, 23 cases were negative for EBER. 15 cases were positive for c-myc gene signal, and 2 cases were positive for c-myc gene signal. 2 cases were positive for c-myc gene signal, 1 case was positive for EBER gene, and 1 case was positive for c-myc gene breakage in 50%).Raji cell line. 3Compared with EBER negative BL cases and tonsil tissues, miR-127 expression of EBER positive BL cases was up-regulated. The difference was statistically significant. Compared with Ramos cell line and Ramos cell line, miR-127 expression was up-regulated in Ramos cell line. Compared with EBER negative BL cases, the expression of mmiR-155 was up-regulated in EBER positive BL cases, but there was no significant difference between them and tonsil tissues. The difference was statistically significant. In Raji cell line, the expression of miR-155 up-regulated in EBV positive BL was significantly lower than that in EBV negative BL(0.504 卤1.214) and in tonsils, the expression of miR-200a was significantly lower than that of Ramos and Ramos. There was no significant difference between EBV positive BL and c-myc gene expression. Conclusion: The expression of miR-127 miR-155 and miR-200a in sporadic BL of children with different EBV infection status was different. The high expression of miR-127 miR-155 and the low expression of miR-200a are closely related to the pathogenesis of sporadic BL in children and the infection of EBV, which may play the role of oncogene / tumor suppressor gene.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R733.1

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