本文選題：天皰瘡 + 差異表達(dá)��； 參考：《西南醫(yī)科大學(xué)》2017年碩士論文

【摘要】：尋常型天皰瘡(PV)是慢性發(fā)皰性皮膚病中的一種常見類型,其病因至今尚未明確,通常被認(rèn)為是一種自身免疫反應(yīng)[1]。當(dāng)前對其治療多采用糖皮質(zhì)激素,長期應(yīng)用存在副作用問題甚至濫用可危及生命。因而進(jìn)一步探究PV的發(fā)病機(jī)制,尋找潛在的關(guān)鍵性生物學(xué)調(diào)控位點(diǎn)顯得尤為迫切。長鏈非編碼RNA(LncRNA)已被證實可以在表觀遺傳、轉(zhuǎn)錄和轉(zhuǎn)錄后等水平調(diào)節(jié)基因的表達(dá),與很多疾病的發(fā)生、發(fā)展有著密切的聯(lián)系。有關(guān)LncRNA與PV的研究國內(nèi)外未見報道。目的:本研究擬應(yīng)用基因芯片技術(shù)對PV患者PBMC中LncRNA和mRNA的差異性表達(dá)情況進(jìn)行檢測,并進(jìn)行初步的生物信息學(xué)分析。方法:采用基因芯片技術(shù)篩選PV患者和正常人PBMC中LncRNA和mRNA的差異性表達(dá),并采用qRT-PCR的方法進(jìn)行驗證;對差異性表達(dá)的LncRNA進(jìn)行Cis和Trans靶基因預(yù)測,間接推斷Lnc RNA的生物學(xué)功能;對差異性表達(dá)的m RNA進(jìn)行GO和KEGG富集分析,進(jìn)一步了解其作用機(jī)制及生物學(xué)功能;將LncRNA靶基因和差異性mRNA功能富集的結(jié)果進(jìn)行比較分析,找尋PV發(fā)生發(fā)展的可能作用機(jī)制或信號通路,為后續(xù)LncRNA功能驗證打下基礎(chǔ)。結(jié)果:共發(fā)現(xiàn)LncRNA 40343個,其中差異性表達(dá)的LncRNA219個,上調(diào)142個,占總數(shù)的0.35198%;下調(diào)77個,占總數(shù)的0.19086%;共發(fā)現(xiàn)mRNA 21422個,其中差異性表達(dá)的mRNA 74個,上調(diào)34個,占總數(shù)的0.15871%;下調(diào)40個,占總數(shù)的0.18672%;選取其中4個LncRNA進(jìn)行qRT-PCR驗證,結(jié)果與基因芯片檢測結(jié)果相符,證明芯片檢測是準(zhǔn)確、可靠的。共發(fā)現(xiàn)靶基因1484個,其中137個cis靶基因,1347個Trans靶基因。所得靶基因主要富集在以下以下生物學(xué)過程:p53信號轉(zhuǎn)導(dǎo)通路/細(xì)胞內(nèi)吞/溶酶體/細(xì)胞凋亡/氨基酸的代謝及降解/脂肪酸降解/核糖體的功能及調(diào)控/RNA轉(zhuǎn)運(yùn)/Toll樣受體信號通路/T細(xì)胞受體信號通路/前列腺癌/過氧化物酶體/子宮內(nèi)膜癌/慢性骨髓性白血病/B細(xì)胞受體信號通路/急性骨髓性白血病等;差異性表達(dá)的mRNA主要參與了調(diào)節(jié)PPARs信號傳導(dǎo),正向調(diào)節(jié)趨化因子分泌,正向調(diào)節(jié)耐受誘導(dǎo),肝素的生物合成及代謝過程等生物學(xué)過程和核糖體、Jak-STAT信號通路、造血細(xì)胞譜系、細(xì)胞因子受體(Cytokine receptors)相互作用等信號通路。發(fā)現(xiàn)主要通路里的三個mRNA:RPL21、IDO1、CCR3,即是LncRNA的靶基因,也是差異表達(dá)的mRNA。結(jié)論:PV患者和正常人對比,外周血PBMC中存在LncRNA的差異性表達(dá),qRT-PCR進(jìn)一步驗證了芯片結(jié)果的的準(zhǔn)確性。LncRNA異常表達(dá)在PV的發(fā)生、發(fā)展中可能扮演了重要角色。這些LncRNA很有可能是通過Cis或者Trans調(diào)控靶基因,進(jìn)而影響了核糖體、Jak-STAT信號通路、造血細(xì)胞譜系、細(xì)胞因子受體相互作用等信號通路,參與了PV的發(fā)生、發(fā)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.
【學(xué)位授予單位】：西南醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R758.66

【參考文獻(xiàn)】

相關(guān)期刊論文 前8條

1 高謙;周暉;李文靜;;超保守區(qū)非編碼RNA在銀屑病外周血單個核細(xì)胞中差異表達(dá)的初步研究[J];皮膚性病診療學(xué)雜志;2014年05期

2 陳蘭芳;唐元家;沈南;;NEAT1與系統(tǒng)性紅斑狼瘡發(fā)病的相關(guān)研究[J];現(xiàn)代免疫學(xué);2014年05期

3 王慧叢;付萍;;CD_4~+T淋巴細(xì)胞與尋常型天皰瘡相關(guān)性研究進(jìn)展[J];皮膚病與性病;2014年01期

4 章杰;李文芳;王芳;楊紅華;萬蕓;劉偉;;長鏈非編碼RNA H19對瘢痕疙瘩成纖維細(xì)胞增殖的影響[J];實用中西醫(yī)結(jié)合臨床;2013年09期

5 彭武建;王紅蕾;歐陽昕;戴勇;;系統(tǒng)性紅斑狼瘡患者長鏈非編碼RNA差異性表達(dá)研究[J];中國現(xiàn)代醫(yī)學(xué)雜志;2012年11期

6 許文嶸;范衛(wèi)新;;Micro RNA與皮膚[J];臨床皮膚科雜志;2010年07期

7 葉帆;吳舜澤;逯元堂;李鍵;;水資源消耗和COD排放的行業(yè)特征分析[J];環(huán)境與可持續(xù)發(fā)展;2008年05期

8 尚建基;;天皰瘡58例臨床分析[J];中國中西醫(yī)結(jié)合皮膚性病學(xué)雜志;2007年03期

相關(guān)碩士學(xué)位論文 前5條

1 韓俊嶺;hTrm61基因?qū)Π螂装?637細(xì)胞侵襲性的影響及基因芯片差異基因的初步探討[D];鄭州大學(xué);2015年

2 唐莉華;lncRNA HOTAIR、MDA9蛋白及GRP78蛋白在惡性黑色素瘤侵襲和轉(zhuǎn)移中的研究[D];安徽醫(yī)科大學(xué);2014年

3 劉悅;胃癌基因表達(dá)譜特征分析及分子標(biāo)記物的篩選[D];上海交通大學(xué);2014年

4 曹忠波;改進(jìn)的雙聚類算法在癌癥基因芯片數(shù)據(jù)中的應(yīng)用[D];吉林大學(xué);2009年

5 黃躍峰;超水稻雜交基因研究和數(shù)據(jù)庫構(gòu)建[D];吉林大學(xué);2008年

，

本文編號：1878608