本文選題：microRNA + 鼻咽癌　； 參考：《復(fù)旦大學(xué)》2010年碩士論文

【摘要】：microRNA是最近研究發(fā)現(xiàn)的一類長(zhǎng)度約為19～25個(gè)核苷酸的非編碼單鏈小RNA分子,其通過與靶基因的3’端非翻譯區(qū)互補(bǔ)配對(duì)結(jié)合的方法,從而調(diào)控內(nèi)源基因的表達(dá)。miRNA作為生物體生長(zhǎng)發(fā)育的重要調(diào)控基因,其主要研究方向是探明miRNAs與疾病的關(guān)系,尤其是miRNA與腫瘤的關(guān)系。 雖然近年來對(duì)miRNA的研究越來越廣泛,但多集中于肺癌、前列腺癌、乳腺癌、結(jié)腸直腸癌等較為常見和廣泛的癌癥。因?yàn)楸茄拾┑牡貐^(qū)分布極不平衡,具有明顯的種族聚集性和地域性,因此目前國外對(duì)其的研究還不是很深入。鼻咽癌是一種上皮源性惡性腫瘤,絕大多數(shù)屬于低分化鱗狀細(xì)胞癌,惡性程度高,早期易發(fā)生淋巴結(jié)和全身遠(yuǎn)處轉(zhuǎn)移,具有相當(dāng)高的死亡率。據(jù)世界衛(wèi)生組織估計(jì),鼻咽癌80%發(fā)生于中國,尤其高發(fā)于中國廣西及廣東地區(qū)。因此我們意在探索性的研究miRNA在鼻咽癌發(fā)生發(fā)展中的調(diào)控作用。 我們利用Illumina microRNA表達(dá)譜芯片,對(duì)8例鼻咽咽癌組織樣本和4例正常組織樣本進(jìn)行了miRNA表達(dá)譜分析。運(yùn)用Illumina BeadStudio軟件讀取芯片數(shù)據(jù),完成芯片數(shù)據(jù)歸一化及篩選修補(bǔ)處理后,利用SAM軟件及Cluster軟件進(jìn)行差異基因分析及聚類分析。 我們篩選出了41條在鼻咽癌組織和正常組織中差異表達(dá)的miRNA,其中包括6條上調(diào)基因(miR-135b, miR-18a, miR-106b等)和35條下調(diào)基因(miR-34b, miR-34c, miR-10b, let-7, miR-449, miR-497等)。為了驗(yàn)證芯片結(jié)果的可靠性,我們應(yīng)用定量RT-PCR的方法檢測(cè)了差異表達(dá)基因miR-34、let-7g以及非差異表達(dá)基因miR-30c的表達(dá)情況,RT-PCR結(jié)果與芯片結(jié)果吻合。同時(shí),我們還發(fā)現(xiàn)這些差異表達(dá)miRNA的聚類結(jié)果能夠較好的與病人的臨床分期聯(lián)系起來。 我們通過Targetscan數(shù)據(jù)庫,對(duì)差異表達(dá)miRNA的靶基因進(jìn)行預(yù)測(cè),并將預(yù)測(cè)結(jié)果輸入GenMAPP軟件進(jìn)行生物學(xué)通路分析。在通路分析中,大部分靶基因富集度高的通路都涉及到信號(hào)轉(zhuǎn)導(dǎo)及與癌癥發(fā)生相關(guān)的通路,例如EGFR-1信號(hào)通路,MAPK信號(hào)通路等等。其中,小GTP酶介導(dǎo)的信號(hào)轉(zhuǎn)導(dǎo)通路最為顯著。同時(shí),Wnt信號(hào)通路的顯著性,則與我們實(shí)驗(yàn)室過去在鼻咽癌cDNA表達(dá)譜的研究結(jié)果相吻合。 繼續(xù)深入研究這些差異miRNA在生物學(xué)通路中的調(diào)控作用,以及尋找與鼻咽癌發(fā)生發(fā)展進(jìn)程相關(guān)的臨床診斷標(biāo)記物,則是我們未來的努力的方向。
[Abstract]:MicroRNA is a class of non-coding single-stranded small RNA molecules with a length of about 19- 25 nucleotides, which are found by complementary pairing with the 3'untranslated region of the target gene. Therefore, the expression of endogenous genes is regulated by .miRNA as an important regulatory gene for the growth and development of organism. Its main research direction is to find out the relationship between miRNAs and disease, especially the relationship between miRNA and tumor. Although the research on miRNA has been more and more extensive in recent years, it mainly focuses on the common and widespread cancers such as lung cancer, prostate cancer, breast cancer, colorectal cancer and so on. Because the regional distribution of nasopharyngeal carcinoma is very unbalanced and has obvious ethnic aggregation and regionality, the research on nasopharyngeal carcinoma at present is not very thorough. Nasopharyngeal carcinoma (NPC) is an epithelial-derived malignant tumor, most of which belong to poorly differentiated squamous cell carcinoma. The World Health Organization estimates that 80% of nasopharyngeal carcinomas occur in China, especially in Guangxi and Guangdong. Therefore, we intend to explore the regulatory role of miRNA in the carcinogenesis and development of nasopharyngeal carcinoma. The miRNA expression profiles of 8 nasopharyngeal carcinoma tissues and 4 normal tissues were analyzed by Illumina microRNA microarray. Illumina BeadStudio software is used to read chip data, and after normalization of chip data and screening repair, differential gene analysis and clustering analysis are carried out by SAM software and Cluster software. We screened 41 differentially expressed miRNAs in nasopharyngeal carcinoma and normal tissues, including 6 up-regulated genes miR-135b, miR-18a, miR-106b) and 35 down-regulated genes miR-34b, miR-34c, miR-10b, let-7, miR-449, miR-497 and so on. In order to verify the reliability of the microarray results, the differential expression gene miR-34let-7g and the non-differentially expressed gene miR-30c were detected by quantitative RT-PCR. The results of RT-PCR were consistent with those of the microarray. At the same time, we also found that the clustering results of these differentially expressed miRNA could be well correlated with the clinical stages of the patients. The target genes of differentially expressed miRNA were predicted by Targetscan database, and the predicted results were input into GenMAPP software for biological pathway analysis. In pathway analysis, most of the pathways with high target gene enrichment are involved in signal transduction and carcinogenesis related pathways, such as EGFR-1 signaling pathway, MAPK signaling pathway and so on. Among them, the signal transduction pathway mediated by small GTP enzyme was the most significant. At the same time, the significance of Wnt signaling pathway was consistent with the previous results of our laboratory studies on cDNA expression profiles in nasopharyngeal carcinoma (NPC). Further study on the regulatory role of miRNA in biological pathways and identification of clinical diagnostic markers related to the progression of nasopharyngeal carcinoma are the directions of our future efforts.
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R739.63

【參考文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

1 魏慶;cDNA芯片在癌癥研究中的應(yīng)用[D];復(fù)旦大學(xué);2006年

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本文編號(hào)：1869649