發(fā)布時間：2018-03-31 20:25

  本文選題：表皮生長因子受體　切入點：長鏈非編碼RNA　出處：《青島大學(xué)》2017年碩士論文

【摘要】：目的:肺癌是常見的惡性腫瘤之一,隨著醫(yī)學(xué)的發(fā)展,手術(shù)、放化療、靶向治療及免疫治療等治療模式都取得了巨大的進步,但肺癌仍是全球癌癥死亡的首要原因。按照不同的組織學(xué)的特點,肺癌大體可以分為兩種,小細胞型及非小細胞型,其中大部分屬為非小細胞型。研究發(fā)現(xiàn)在非小細胞肺癌患者中大約40%-80%能夠過度表達EGFR,以EGFR為靶點的治療模式較野生型和傳統(tǒng)化療能明顯改善突變患者的生存,已成為NSCLC治療的新熱點。在龐大的人類基因組當(dāng)中,絕大多數(shù)的基因無法進行轉(zhuǎn)錄,而余下的極少數(shù)基因才能夠進行轉(zhuǎn)錄,這些基因當(dāng)中98%以上的基因?qū)儆诜蔷幋aRNA(non-coding RNA,nc RNA)。按照鏈的長度大小可將非編碼RNA分為短鏈、中鏈和長鏈非編碼lncRNA(long non-coding RNA)。lncRNA至少有200個核苷酸,是一種不編碼蛋白質(zhì)的RNA。大量研究顯示,lncRNA的異常表達對腫瘤的形成和演進有重要的作用。另外,研究發(fā)現(xiàn)lncRNA與肝癌、肺癌、乳腺癌、胃癌、結(jié)直腸癌等多種腫瘤關(guān)系密切。因此,對EGFR突變肺癌的lncRNA表達譜進行生物信息學(xué)方面的研究也為肺癌的研究增添了一個新的切入點。我們的研究目的是利用生物信息學(xué)分析EGFR突變肺癌中差異表達的lncRNA,對篩選的lncRNA進行功能分析。方法:從NCBI公共數(shù)據(jù)平臺GEO下載肺癌基因芯片數(shù)據(jù)GSE31210,共獲得127例EGFR突變肺癌和20例正常組織的基因表達譜數(shù)據(jù)。采用穩(wěn)健多芯片平均標(biāo)準(zhǔn)化(RMA分析)方法對GSE31210表達譜數(shù)據(jù)進行標(biāo)準(zhǔn)化預(yù)處理,預(yù)處理后的結(jié)果用微陣列顯著性分析(SAM)軟件進行分析,找出在EGFR突變肺癌中有顯著表達差異的lncRNA。對收集的19例EGFR突變肺癌組織和癌旁組織標(biāo)本進行組織RNA的提取,經(jīng)熒光定量PCR驗證篩選出的候選基因lncRNA CR749465的差異表達。利用DAVID在線軟件分析候選lncRNACR749465相關(guān)靶基因,進行GO功能分類及pathway通路分析。結(jié)果:以實驗設(shè)定的標(biāo)準(zhǔn)作為差異基因篩選標(biāo)準(zhǔn)(P≤0.05;|(fold change)|≥2),GSE31210芯片篩選的結(jié)果表明,在EGFR突變肺癌組織和正常組織中,lncRNA的表達譜有明顯差異,有127個基因出現(xiàn)差異表達。在篩選所得的差異lncRNA127個中,其中上調(diào)的有96個,下調(diào)的有31個。其中上調(diào)變化最大的是CR749465,有3.26倍的表達變化差異。經(jīng)過熒光定量PCR驗證,發(fā)現(xiàn)候選基因lncRNACR749465基因表達情況與芯片數(shù)據(jù)分析結(jié)果一致。lncRNA CR749465相關(guān)靶基因GO分析顯示CR749465相關(guān)基因可能與血管生成、信號傳導(dǎo)、RAN代謝等有關(guān)。Pathway分析顯示CR749465相關(guān)基因可能主要與神經(jīng)活性的配體-受體相互作用通路、軸突導(dǎo)向通路、細胞粘附分子通路、Rap1信號通路等有關(guān)。結(jié)論:EGFR突變肺癌存在明顯的lncRNA差異性表達.通過GO功能分析及KEGG通路分析發(fā)現(xiàn),CR749465可能在EGFR突變肺癌的發(fā)生和發(fā)展中發(fā)揮重要作用。
[Abstract]:Objective: lung cancer is one of the common malignant tumors, with the development of medicine, surgery, radiotherapy, chemotherapy, targeted treatment and immunotherapy have made great progress. But lung cancer is still the leading cause of cancer death in the world. According to different histological features, lung cancer can be divided into two types: small cell type and non small cell type. Most of them were non-small cell type. About 40% to 80% of patients with NSCLC were able to overexpression EGFR. Treatment targeting EGFR could significantly improve the survival of mutant patients compared with wild-type and conventional chemotherapy. Has become a new hotspot in NSCLC therapy. In the vast human genome, the vast majority of genes cannot be transcribed, while the few remaining genes can be transcribed. More than 98% of these genes belong to non-coding RNA(non-coding RNAs or nc-RNAs. According to the length of the chain, the non-coding RNA can be divided into short strands, and there are at least 200 nucleotides in the medium-chain and long-chain non-coding lncRNA(long non-coding RNA).lncRNA. A large number of studies have shown that abnormal expression of LncRNA plays an important role in tumor formation and progression. In addition, lncRNA has been found to be associated with liver cancer, lung cancer, breast cancer and gastric cancer. Colorectal cancer and many other tumors are closely related. Therefore, The bioinformatics study of lncRNA expression profile of EGFR mutant lung cancer also provides a new entry point for the study of lung cancer. Our aim is to analyze the differential expression of EGFR RNA in EGFR mutant lung cancer by bioinformatics. Functional analysis of screened lncRNA. Methods: from NCBI common data platform GEO download lung cancer gene chip data GSE31210, a total of 127 cases of EGFR mutation lung cancer and 20 cases of normal tissue gene expression profile data, using robust multi-chip average. The GSE31210 expression profile data were preprocessed by the method of standardized RMA analysis. The results of preconditioning were analyzed by microarray significance analysis software (Sam) to find out the significant difference in the expression of LNC NNA in EGFR mutant lung cancer. The RNA was extracted from 19 samples of EGFR mutant lung cancer and paracancerous tissues. The differentially expressed candidate gene lncRNA CR749465 was screened by fluorescence quantitative PCR. The candidate lncRNACR749465 related target genes were analyzed by DAVID online software. Go functional classification and pathway pathway analysis were carried out. Results: the differential gene screening criteria (P 鈮，

本文編號：1692314