寻常型天疱疮LncRNA的差异性表达及生物信息学分析

发布时间：2018-05-12 12:24

本文选题：天疱疮 + 差异表达　；参考：《西南医科大学》2017年硕士论文

【摘要】：寻常型天疱疮(PV)是慢性发疱性皮肤病中的一种常见类型,其病因至今尚未明确,通常被认为是一种自身免疫反应[1]。当前对其治疗多采用糖皮质激素,长期应用存在副作用问题甚至滥用可危及生命。因而进一步探究PV的发病机制,寻找潜在的关键性生物学调控位点显得尤为迫切。长链非编码RNA(LncRNA)已被证实可以在表观遗传、转录和转录后等水平调节基因的表达,与很多疾病的发生、发展有着密切的联系。有关LncRNA与PV的研究国内外未见报道。目的:本研究拟应用基因芯片技术对PV患者PBMC中LncRNA和mRNA的差异性表达情况进行检测,并进行初步的生物信息学分析。方法:采用基因芯片技术筛选PV患者和正常人PBMC中LncRNA和mRNA的差异性表达,并采用qRT-PCR的方法进行验证;对差异性表达的LncRNA进行Cis和Trans靶基因预测,间接推断Lnc RNA的生物学功能;对差异性表达的m RNA进行GO和KEGG富集分析,进一步了解其作用机制及生物学功能;将LncRNA靶基因和差异性mRNA功能富集的结果进行比较分析,找寻PV发生发展的可能作用机制或信号通路,为后续LncRNA功能验证打下基础。结果:共发现LncRNA 40343个,其中差异性表达的LncRNA219个,上调142个,占总数的0.35198%;下调77个,占总数的0.19086%;共发现mRNA 21422个,其中差异性表达的mRNA 74个,上调34个,占总数的0.15871%;下调40个,占总数的0.18672%;选取其中4个LncRNA进行qRT-PCR验证,结果与基因芯片检测结果相符,证明芯片检测是准确、可靠的。共发现靶基因1484个,其中137个cis靶基因,1347个Trans靶基因。所得靶基因主要富集在以下以下生物学过程:p53信号转导通路/细胞内吞/溶酶体/细胞凋亡/氨基酸的代谢及降解/脂肪酸降解/核糖体的功能及调控/RNA转运/Toll样受体信号通路/T细胞受体信号通路/前列腺癌/过氧化物酶体/子宫内膜癌/慢性骨髓性白血病/B细胞受体信号通路/急性骨髓性白血病等;差异性表达的mRNA主要参与了调节PPARs信号传导,正向调节趋化因子分泌,正向调节耐受诱导,肝素的生物合成及代谢过程等生物学过程和核糖体、Jak-STAT信号通路、造血细胞谱系、细胞因子受体(Cytokine receptors)相互作用等信号通路。发现主要通路里的三个mRNA:RPL21、IDO1、CCR3,即是LncRNA的靶基因,也是差异表达的mRNA。结论:PV患者和正常人对比,外周血PBMC中存在LncRNA的差异性表达,qRT-PCR进一步验证了芯片结果的的准确性。LncRNA异常表达在PV的发生、发展中可能扮演了重要角色。这些LncRNA很有可能是通过Cis或者Trans调控靶基因,进而影响了核糖体、Jak-STAT信号通路、造血细胞谱系、细胞因子受体相互作用等信号通路,参与了PV的发生、发展。
[Abstract]:Pemphigus vulgaris (PVV) is a common type of chronic bullous dermatosis. The etiology of PVV is still unknown and is generally considered to be an autoimmune response [1]. At present, glucocorticoids are often used in the treatment of glucocorticoid. Long-term application has side effects and even misuse can be life-threatening. Therefore, it is urgent to further explore the pathogenesis of PV and to find potential key biological regulatory sites. LncRNAs have been proved to regulate gene expression at epigenetic, transcriptional and post-transcriptional levels, which is closely related to the occurrence and development of many diseases. The research on LncRNA and PV has not been reported at home and abroad. Objective: to detect the differential expression of LncRNA and mRNA in PBMC of PV patients by using gene chip technique and to carry out preliminary bioinformatics analysis. Methods: the differential expression of LncRNA and mRNA in PBMC of PV patients and normal subjects was screened by gene chip technique and verified by qRT-PCR method, Cis and Trans target genes were predicted by Cis and Trans target genes were predicted, and the biological function of Lnc RNA was inferred indirectly. The m RNA expressed differently was enriched by go and KEGG, and its mechanism and biological function were further understood. The results of LncRNA target gene and differential mRNA function enrichment were compared and analyzed. To find out the possible mechanism or signal pathway of PV, and to lay a foundation for the subsequent functional verification of LncRNA. Results: a total of 40343 LncRNA were found, of which 142 were up-regulated (0.35198), 77 were down-regulated (0.190868), and mRNA 21422 were found, of which 74 were differentially expressed mRNA, 34 were up-regulated (0.15871), and 40 were down-regulated. 4 of them were selected for qRT-PCR verification, the results were consistent with the results of gene chip detection, which proved that the chip detection was accurate and reliable. A total of 1484 target genes were identified, of which 137 were cis target genes and 1347 Trans target genes. The resulting target genes are mainly enriched in the following biological processes: signal transduction pathway / endocytosis / lysosome / apoptosis / amino acid metabolism and degradation / fatty acid degradation / ribosomal function and regulation of rRNA transport r-Toll T-cell receptor signaling pathway / prostate cancer / peroxisome / endometrial carcinoma / chronic myeloid leukemia / acute myeloid leukemia. Differentially expressed mRNA is involved in the regulation of PPARs signal transduction, forward regulation of chemokine secretion, forward regulation of tolerance induction, biosynthesis and metabolism of heparin, ribosomal Jak-STAT signaling pathway, hematopoietic lineage, etc. Cytokine receptor Cytokine receptor interaction and other signaling pathways. Three mRNAs: RPL21, IDO1, CCR3, the target gene of LncRNA, were found to be differentially expressed mRNAs. Conclusion the existence of differential expression of LncRNA in peripheral blood of PBMC patients and normal controls further verifies the accuracy of microarray results. Abnormal expression of LncRNA may play an important role in the pathogenesis of PV and may play an important role in the development of PV. These LncRNA may regulate target genes through Cis or Trans, and then affect the signal pathways such as ribosomal Jak-STAT signaling pathway, hematopoietic lineage, cytokine receptor interaction and participate in the occurrence and development of PV.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R758.66

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