银屑病皮损与正常皮肤组织miRNA表达差异分析

发布时间：2018-10-11 15:13

【摘要】：目的:利用miRNA芯片技术分析银屑病皮损与正常皮肤组织之间miRNA表达差异,同时对差异表达的miRNA所调控的靶基因进行预测和分子功能分析,以及如何参与银屑病发生等方面进行探讨,旨在进一步阐述本病发病机制的基础上为临床的靶向治疗提供依据。方法:本研究选择寻常型银屑病进行期3例,正常对照组3例,分别取皮损组织和正常皮肤组织进行总RNA提取,质量、纯度鉴定,符合Affymetrix miRNA芯片技术要求后按照miRNA芯片标准试实验流程进行荧光标记,杂交,然后利用Affymetrix GeneChip Scanner 3000激光共聚焦扫描仪对杂交结果进行图像扫描,配合 Affymetrix GeneChip Operating Software Version1.4 数据处理软件读取、处理数据;按照q-value(%)≤5,同时差异倍数(Fold Change)控制在2倍以上的标准筛选差异表达的miRNA作为分析数据的来源。对差异表达的miRNA进行靶基因预测和功能分析分别利用Human Target scan 5.1数据库和MAS系统Pathway和GO分类数据库。结果:①3例寻常型银屑病患者皮损组织与3例正常人皮肤组织相比较,差异表达的miRNA有5条,其中Hsa-miR-1308,表达上调;Hsa-miR-27a,Hsa-miR-199a-3p,Hsa-miR-199b-3p,Hsa-miR-181a,表达均下调,5 条差异表达的miRNA序列长度均集中在18～23个核苷酸之间。②Human Target scan5.1数据库对5条差异表达的miRNA所负调控的靶基因进行预测,数目达到2168条;利用MAS系统Pathway和GO分类数据库分析2168条靶基因,结果显示,这些靶基因主要涉及多种信号通路的传导,角质形成细胞的增殖与分化,免疫细胞增殖与调控等生物学过程。结论:银屑病是一种遗传基因异常性疾病,转录水平上的miRNA可靶向负调控大量mRNA,而这些被调控的mRNA之间又存在错综复杂的相互影响和作用,共同形成网络式的调节结构通过多种途径来参与银屑病的发生。
[Abstract]:Objective: to analyze the difference of miRNA expression between psoriatic lesions and normal skin tissues by using miRNA microarray, and to predict and analyze the molecular function of target genes regulated by differentially expressed miRNA. And how to participate in the occurrence of psoriasis were discussed in order to further explain the pathogenesis of psoriasis on the basis of clinical targeted therapy. Methods: in this study, 3 cases of psoriasis vulgaris and 3 cases of normal control group were selected for total RNA extraction, quality and purity identification. According to the technical requirements of Affymetrix miRNA chip, fluorescence labeling and hybridization were carried out according to the experimental procedure of miRNA chip standard, and then the result of hybridization was scanned by Affymetrix GeneChip Scanner 3000 laser confocal scanner. The Affymetrix GeneChip Operating Software Version1.4 data processing software is used to read and process the data, and according to q-value (%) 鈮，

本文编号：2264524