高通量测序研究卵巢子宫内膜异位症中异常lncRNA表达
发布时间：2018-12-30 17:11
【摘要】：子宫内膜异位症(endometriosis, EMs),是一种常见的妇科疾病,育龄期妇女发病率高达10-15%,病灶可侵犯全身任何组织,卵巢子宫内膜异位症是其常见的类型之一。尽管其具体发病机制不明,但一般认为内异症是一种多基因导致的疾病。研究表明长链非编码RNA (long non-coding RNA, lncRNA)的异常表达与多种疾病相关,目前与子宫内膜异位症相关的lncRNA研究还较少。高通量测序技术是研究lncRNA的一种重要方法,国内外目前尚无高通量测序对于子宫内膜异位症的研究,本实验首次采用高通量测序技术对异位内膜(ectopic endometrium, EC)、在位内膜(eutopic endometrium,EU )及正常内膜(normal endometrium, N)组织进行检测,拟为内异症相关的lncRNA的后续研究提供候选基因库,希望有助于进一步揭示lncRNA在内异症发生发展过程中的具体作用机制,及为后期寻找新的靶向治疗药物提供实验基础。第一部分:IncRNA在卵巢子宫内膜异位症中的表达谱分析目的:高通量测序技术筛选卵巢子宫内膜异位症中异位内膜、在位内膜和正常内膜组织间差异表达的lncRNAs,并进行初步的功能研究,探寻内异症可能得发病机制。方法:采集8对人异位内膜、在位内膜组织及5例正常内膜组织进行高通量测序检测,并用生物信息技术对测序得到的海量信息进行分析,筛选三者间差异表达的lncRNAs,对差异lncRNAs进行功能注释。结果:比较异位内膜、在位内膜及正常内膜组之间差异表达lncRNAs,发现异位内膜与在位内膜之间有683个已知lncRNAs和443个新发现的lncRNAs异常表达;在位内膜与正常内膜之间有5个已知lncRNAs和56个新发现的lncRNAs异常表达;异位内膜与正常内膜之间有603个已知lncRNAs和408个新发现的lncRNAs异常表达。对其进行生物信息学分析发现,异常表达的lncRNAs参与多种内异症的分子功能、细胞组分和生物过程中。结论:本研究首次采用了高通量测序技术对正常内膜、在位内膜及异位内膜组织进行检测,发现三者间有大量异常表达的lncRNAs,这些lncRNAs可能在卵巢子宫内膜异位症的发生发展中发挥着重要的作用。第二部分:IncRNAGAPLINC、CTD-2207P18.1 和 RP11-1100L3.8 在卵巢子宫内膜异位症中的表达分析目的:验证 lncRNAGAPLINC、CTD-2207P18.1 和 RP11-1100L3.8 在异位内膜、在位内膜组织及正常内膜组织中的差异表达,并探寻其可能作用途径。方法:提取20对配对的人异位内膜、在位内膜组织及5例正常内膜组织的总RNA,利用实时定量聚合酶链锁反应(quantitative real time PCR,qRT-PCR)方法对3个差异表达的lncRNAs进行检测,2-△△Ct的方法计算不同组织的实时定量数据,组间差异采用t检验,P0.05认为差异有统计学意义。并对这3个异常表达的lncRNAs进行生物信息学分析。结果:lncRNAGAPLINC和CTD-2207P18.1异位内膜中表达水平明显高于在位内膜,;ncRNARP11-1100L3.8在位内膜中表达水平高于正常内膜,三者参与到内异症的多个GO条目的富集及KEGG通路。结论:qRT-PCR结果分析与高通量测序一致。异位内膜、在位内膜中;ncRNA GAPLINC、CTD-2207P18.1、RP11-1100L3.8存在差异表达,这三者可能参与卵巢子宫内膜异位症的的发生发展。
[Abstract]:Endometriosis (EMs) is a common gynecological disease. The incidence of women in the childbearing potential is up to 10-15%. Although the specific pathogenesis is unknown, it is generally considered to be a multi-gene-induced disease. The research shows that the abnormal expression of long-chain non-coding RNA (lncRNA) is associated with a variety of diseases, and the research on the ncRNA associated with endometriosis is still less. high-throughput sequencing technology is an important method to study lncRNA, and there is no high-throughput sequencing at home and abroad for endometriosis. N) The tissue was tested to provide a candidate gene bank for the follow-up study of the lncRNA associated with the endosiosis, and it was hoped to help to further reveal the specific mechanism of the lncRNA in the development of the isodisease, and to provide an experimental basis for finding new targeted therapeutic drugs in the later stage. The first part: The purpose of the expression of IncRNA in the diagnosis of ovarian endometriosis: the high-throughput sequencing technique is used to screen the expression of the ncRNAs in the ectopic endometrium, the in-place and the normal endometrium of the endometriosis, and to carry out a preliminary functional study. It is possible to explore the possible mechanism of internal heterosis. Methods: 8 pairs of ectopic endometrium, in-place and 5 normal endometrium were collected for high-throughput sequencing, and the mass information obtained by sequencing was analyzed by using the biological information technology. Results: The abnormal expression of lncRNAs was found between the ectopic endometrium, the in-place and the normal endometrium group, and there were 683 known lncRNAs and 443 newly discovered lncRNAs, and there were 5 known lncRNAs and 56 newly discovered lncRNAs in the endometrium and the normal endometrium. There were 603 known lncRNAs and 408 newly discovered lncRNAs abnormal expression between the ectopic endometrium and the normal endometrium. Bioinformatics analysis shows that the abnormal expression of lncRNAs is involved in the molecular function, cellular components and biological processes of a variety of endothers. Conclusion: The first time in this study, high-throughput sequencing technology was used to detect the normal endometrium, in-place and ectopic endometrium, and it was found that there was a large number of abnormal expression of lncRNAs, which could play an important role in the development of endometriosis. The second part: The purpose of the expression and analysis of IncRNA GAPLINC, CTD-2207P18.1 and RP11-1100L3. 8 in the diagnosis of ovarian endometriosis: to verify the differential expression of lncRNAGAPLINC, CTD-2207P18.1 and RP11-1100L3. 8 in the ectopic endometrium, in-place and in the normal endometrium, and to explore the possible ways of action. Methods: The total RNA of 20 pairs of human heterotopic endometrium, in-place and 5 normal endometrium was extracted, and 3 differentially expressed lncRNAs were detected by the method of real-time quantitative polymerase chain reaction (qRT-PCR). The real-time quantitative data of different tissues was calculated by the method of 2-Thiophans. The difference between the groups was t-test, and the difference was considered to be of statistical significance. The bioinformatics analysis of the three abnormal expression of lncRNAs was carried out. Results: The expression level of ncRNAGAPLINC and CTD-2207P18. 1 was significantly higher than that in the in-place, and the expression of ncRNARP11-1100L3. 8 was higher than that in the normal endometrium. Conclusion: The results of qRT-PCR are consistent with high-throughput sequencing. The differential expression of ncRNA GAPLINC, CTD-2207P18.1, RP11-1100L3. 8 may be involved in the development of endometriosis.
【学位授予单位】：中国人民解放军医学院
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R711.71
本文编号：2395876
[Abstract]:Endometriosis (EMs) is a common gynecological disease. The incidence of women in the childbearing potential is up to 10-15%. Although the specific pathogenesis is unknown, it is generally considered to be a multi-gene-induced disease. The research shows that the abnormal expression of long-chain non-coding RNA (lncRNA) is associated with a variety of diseases, and the research on the ncRNA associated with endometriosis is still less. high-throughput sequencing technology is an important method to study lncRNA, and there is no high-throughput sequencing at home and abroad for endometriosis. N) The tissue was tested to provide a candidate gene bank for the follow-up study of the lncRNA associated with the endosiosis, and it was hoped to help to further reveal the specific mechanism of the lncRNA in the development of the isodisease, and to provide an experimental basis for finding new targeted therapeutic drugs in the later stage. The first part: The purpose of the expression of IncRNA in the diagnosis of ovarian endometriosis: the high-throughput sequencing technique is used to screen the expression of the ncRNAs in the ectopic endometrium, the in-place and the normal endometrium of the endometriosis, and to carry out a preliminary functional study. It is possible to explore the possible mechanism of internal heterosis. Methods: 8 pairs of ectopic endometrium, in-place and 5 normal endometrium were collected for high-throughput sequencing, and the mass information obtained by sequencing was analyzed by using the biological information technology. Results: The abnormal expression of lncRNAs was found between the ectopic endometrium, the in-place and the normal endometrium group, and there were 683 known lncRNAs and 443 newly discovered lncRNAs, and there were 5 known lncRNAs and 56 newly discovered lncRNAs in the endometrium and the normal endometrium. There were 603 known lncRNAs and 408 newly discovered lncRNAs abnormal expression between the ectopic endometrium and the normal endometrium. Bioinformatics analysis shows that the abnormal expression of lncRNAs is involved in the molecular function, cellular components and biological processes of a variety of endothers. Conclusion: The first time in this study, high-throughput sequencing technology was used to detect the normal endometrium, in-place and ectopic endometrium, and it was found that there was a large number of abnormal expression of lncRNAs, which could play an important role in the development of endometriosis. The second part: The purpose of the expression and analysis of IncRNA GAPLINC, CTD-2207P18.1 and RP11-1100L3. 8 in the diagnosis of ovarian endometriosis: to verify the differential expression of lncRNAGAPLINC, CTD-2207P18.1 and RP11-1100L3. 8 in the ectopic endometrium, in-place and in the normal endometrium, and to explore the possible ways of action. Methods: The total RNA of 20 pairs of human heterotopic endometrium, in-place and 5 normal endometrium was extracted, and 3 differentially expressed lncRNAs were detected by the method of real-time quantitative polymerase chain reaction (qRT-PCR). The real-time quantitative data of different tissues was calculated by the method of 2-Thiophans. The difference between the groups was t-test, and the difference was considered to be of statistical significance. The bioinformatics analysis of the three abnormal expression of lncRNAs was carried out. Results: The expression level of ncRNAGAPLINC and CTD-2207P18. 1 was significantly higher than that in the in-place, and the expression of ncRNARP11-1100L3. 8 was higher than that in the normal endometrium. Conclusion: The results of qRT-PCR are consistent with high-throughput sequencing. The differential expression of ncRNA GAPLINC, CTD-2207P18.1, RP11-1100L3. 8 may be involved in the development of endometriosis.
【学位授予单位】：中国人民解放军医学院
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R711.71
【参考文献】
相关期刊论文 前6条
1 刘潺;石雅宁;朱能;杜可;倪亚光;廖端芳;覃丽;;长链非编码RNA类固醇受体RNA激活子在乳腺癌中的表达及作用机制研究状况[J];中国临床药理学杂志;2016年12期
2 张素洁;胡毅;马俊勋;;长链非编码RNA检测在肿瘤诊断及预后评估中的应用综述[J];解放军医学院学报;2016年08期
3 张琛;关菁;骆健俊;吴薇;叶雪;程洪艳;马瑞琼;崔恒;昌晓红;;长链非编码RNA CHL1-AS2在卵巢子宫内膜异位症中的表达及意义初探[J];中国妇产科临床杂志;2016年01期
4 邹瑞;林颖;欧阳可雄;朴正国;廖贵清;;H19基因在肿瘤发生发展中的作用研究进展[J];实用医学杂志;2015年19期
5 朱晓雁;王迪;董梅;;长链非编码RNA参与固有免疫应答调控的研究进展[J];解放军医学院学报;2015年12期
6 王俊豪;阮钰;李蓉;;印迹基因H19和Igf2在子宫内膜异位症患者子宫内膜组织中的异常表达及其意义[J];生殖医学杂志;2013年07期
，本文编号：2395876
本文链接：https://www.wllwen.com/yixuelunwen/fuchankeerkelunwen/2395876.html
最近更新
教材专著
