microrna论文：子宫内膜样腺癌组织中microrna差异表达谱分析及其靶基因预测的研究

microRNA 论文：子宫内膜样腺癌组织中 microRNA 差异表 达谱分析及其靶基因预测的研究 【中文摘要】与子宫内膜癌是累及女性生殖系统最常见的恶性 肿瘤之一,在很多欧美国家,其发病率居女性生殖系统癌症的首位。 而在我国,子宫内膜癌的发病率也仅次于子宫颈癌,居女性生殖系统 癌症的第二位,且有逐年上升趋势。子宫内膜癌中有 85%-90%为型 子宫内膜样腺癌(雌激素依赖型),严重危害女性健康。目前认为子宫 内膜癌的发生可能与长期持续的雌激素刺激或者肥胖、高血压、糖 尿病、不孕等因素相关,但其机制尚不明确。因此对子宫内膜癌尤其 是内膜样腺癌发病机制和新诊断方法的探讨与研究十分必要。 microRNA(MicroRNA)是一种由 22-25 个核苷酸组成的非编码单链小 RNA,目前认为在哺乳动物体内,microRNA 与靶基因作用,使靶基因的 mRNA 在转录后的翻译过程中被抑制或直接降解,导致靶基因的蛋白 表达水平降低,从而间接参与到包括生长发育、造血、器官形成、细 胞增殖、凋亡乃至肿瘤发生的过程中。近些年来,很多研究围绕着 microRNA 与各种肿瘤间的关系展开,而针对子宫内膜样腺癌中 microRNA 变化的研究目前尚不多。本研究通过筛查子宫内膜样腺癌 组织与正常对照组子宫内膜组织中 microRNA 的表达差异,同时结合 已有资料对表达差异明显的 microRNA 进行靶基因预测,并使用微阵 预测分析(PAM)工具初步预测一组可以用于区分不同病理分化子宫内 膜样腺癌的 microRNA。进而从分子学水平探讨子宫内膜样腺癌的发 病机制。方法：1.使用丹麦 ExiqonTM 公司 microRNA 芯片检测出子 宫内膜样腺癌组织和正常内膜组织 microRNA 表达谱,筛选出表达差 异明显的 microRNA。2.挑选出上调及下调明显的 microRNA 各 1 例 使用实时定量逆转录 PCR (real time RT-PCR)进行验证。3.结合以 往研究中与子宫内膜样腺癌及子宫内膜癌关系密切的基因结果,使用 三种不同的预测软件对本实验中出现明显差异表达的 microRNA 进行 靶基因预测,并统合结果。4.使用微阵预测分析工具处理高、中、低 分化子宫内膜样腺癌及正常内膜中表达的 microRNA 数据,初步得出 一组能够区分以上四组样本的 microRNA 标记。结果：1.在芯片覆盖 的 847 条人 microRNA 中,子宫内膜样腺癌组织相较于正常内膜组织 出现 18 个明显上调,31 个明显下调的 microRNA。2.包括 PTEN、FOX 家族等基因在内的 51 种基因被认为是上述差异表达的 microRNA 的 预测靶基因。3.Mir-200c 等 9 种 microRNA 是一组可用于鉴别不同 分化子宫内膜样腺癌的生物标记 microRNA。结论：1.相较于正常内 膜组织,多种 microRNA 在子宫内膜样腺癌中出现明显表达差异。 2.mir-205 等 microRNA 在子宫内膜样腺癌中表现出明显的变化趋势,可 能是影响子宫内膜样腺癌发生发展的关键 microRNA。3.mir- 205、mir-200 家族、mir-183 家族等多种成簇排列的 microRNA 家族 在子宫内膜样腺癌中出现集体性变化。4.PTEN、FOX 等多种基因被 预测为子宫内膜样腺癌中差异表达 microRNA 的靶基因,microRNA 可 能通过调控这些靶基因从而影响子宫内膜样腺癌的发生发展过程。 【英文摘要】Background and :Endometrial carcinoma is one of the most common gynecological malignant tumors. The incidence of endometrial carcinoma in America and European countries ranks first in the gynecologic malignancy. In China, this disease is the second common gynecologic malignancy and ranks after cervical cancer. And its incidence is increasing year by year. As a disease does great harm to womens health, about 85%-90% of all endometrial carcinomas are estrogen-related endometrial cancer (type I). And most of type I endomtetrial cancer cases are endometrioid endometrial adenocarcinoma(EEC). Excessive stimulation of estrogen, obesity, hypertension, diabetes, infertility and other factors are considered related to the tumorigenesis of endometrial cancer currently, but the mechanism is unclear. So, it is meaningful to explore the pathogenesis of endometrial cancer, especially EEC.MicroRNA is a class of non- proteincoding, single-stranded small RNAs of 22-25 nucleotides length. They can regulate multiple target genes. In mammals, target gene expression is repressed by post- transcriptionally. And it takes part in many physiological processes including body growth and development, hematopoiesis, organ formation, cell proliferation, apoptosis even tumorigenesis. In recent years, there are many studies on the relationship between microRNA and tumorgenesis. But few of them referred EEC.In our study, differential expression microRNAs profile was detected from EEC compared with normal endometrium. Then targets of these differential expression microRNAs were predicted through web databases. Prediction analysis for microarray (PAM) was used for the first time to predict a group of microRNA biomarkers. These markers could be used to identify different differentiation of EEC.Methods:MicroRNA microarray was used to detect the differential expression microRNAs, and real time RT-PCR was used to verify the results. Then targets of these microRNAs in EEC were predicted through web databases. PAM was used to predict a group of microRNA biomarkers, which could be used to identify differentiation of EEC.Results:18 obviously up- regulated and 31 obviously down-regulated microRNAs in tumor group were identified.51 genes were predicted as targets of these dysregulated microRNAs. Mir-200c and other 8 microRNAs were considered as members of the biomarkers that could identify differentiation. Many members from same microRNA families (microRNA cluster) changed at the same time in EEC.Conclusion:Several microRNAs were found have special changes in EEC and these microRNAs and their predicted targets might play an important role in EEC. There were a lot of collective microRNAs from the family changed together in EEC in our study. They could co-express the same role in tumorigenesis. The predicted targets of dysregulated microRNA in EEC just like PTEN and FOX might be regulated by microRNA and took part in tumorigenesis. 【关键词】microRNA 子宫内膜样腺癌 microRNA 芯片 靶基因 微阵预测分析 【英文关键词】Micro RNA Endometrioid endometrial adenocarcinoma MicroRNA microarray Target gene Predict