發(fā)布時(shí)間：2018-07-10 11:01

  本文選題：賁門癌 + 長(zhǎng)鏈非編碼RNA　； 參考：《河南科技大學(xué)》2017年碩士論文

【摘要】：目的:利用基因芯片技術(shù)檢測(cè)賁門癌與癌旁組織中長(zhǎng)鏈非編碼RNA(long non-coding RNA,lnc RNA)和微小RNA(microRNA,miRNA)表達(dá)譜;探討賁門癌與癌旁組織中差異性表達(dá)lnc RNA和miRNA之間的相關(guān)性以及異常表達(dá)mi RNA與預(yù)后的關(guān)系。材料與方法:共選取52對(duì)經(jīng)手術(shù)切除的賁門癌與癌旁組織作為研究標(biāo)本,其中15例標(biāo)本用于基因芯片檢測(cè),37例標(biāo)本進(jìn)行后期PCR驗(yàn)證。利用Arraystar Human LncRNA microarray V2.0和miRCURY LNA?micro RNA Array,7th gen-hsa,mmurno兩種芯片來檢測(cè)lncRNA和miRNA的表達(dá)譜,再通過RNA標(biāo)記與雜交、數(shù)據(jù)分析后得出賁門癌與癌旁組織中差異性表達(dá)的lncRNA和miRNA的表達(dá)譜。應(yīng)用生物信息預(yù)測(cè)技術(shù)來分析表達(dá)譜中部分lnc RNA和mi RNA的相關(guān)性,并運(yùn)用qRT-PCR技術(shù)在37對(duì)賁門癌與癌旁組織中驗(yàn)證結(jié)果。運(yùn)用統(tǒng)計(jì)學(xué)分析lncRNA和miRNA兩者之間的表達(dá)相關(guān)性。利用卡方檢驗(yàn)來分析mi RNA與臨床病理特征之間的關(guān)系;運(yùn)用Kaplan-meier生存分析來研究miRNA與生存期的關(guān)系。結(jié)果:15例賁門癌與癌旁組織中2倍以上差異性表達(dá)的lncRNA、miRNA及mRNA的表達(dá)譜:lncRNA共計(jì)2090個(gè);miRNA共計(jì)152。根據(jù)lncRNA表達(dá)譜中差異性表達(dá)倍數(shù)、標(biāo)準(zhǔn)化強(qiáng)度值及l(fā)ncRNA數(shù)據(jù)的注釋,選出兩條lnc RNA:H19和MALAT1。通過生物信息預(yù)測(cè)方法,從賁門癌miRNA表達(dá)譜找到與H19和MALAT1存在表達(dá)相關(guān)性的miRNA共3條,包括:has-mi R-148a-3p、hsa-miR-205-5p及hsa-miR-363-3p。利用qRT-PCR技術(shù)驗(yàn)證一對(duì)預(yù)測(cè)的RNA:H19和has-miR-148a-3p的表達(dá)情況,結(jié)果顯示在37例賁門癌與癌旁組H19呈明顯的過表達(dá)(P=0.0072),mi R-148a-3p呈現(xiàn)明顯的低表達(dá)(P0.0001)。利用非參數(shù)秩和檢驗(yàn)發(fā)現(xiàn)H19與miR-148a-3p的表達(dá)呈現(xiàn)明顯的負(fù)相關(guān)性(P=0.019)。臨床病理特征分析提示低表達(dá)的miRNA與賁門癌淋巴結(jié)轉(zhuǎn)移明顯相關(guān)(P=0.039)。Kaplan-meier生存分析發(fā)現(xiàn)低表達(dá)的miR-148a-3p預(yù)示著較差的賁門癌患者生存期(P=0.022)。結(jié)論:15對(duì)賁門癌與癌旁組織的lnc RNA及miRNA表達(dá)譜存在顯著差異。通過生物信息預(yù)測(cè)和qRT-PCR技術(shù),找到了賁門癌中l(wèi)ncRNA H19和miR-148a-3p存在表達(dá)相關(guān)性。此外,低表達(dá)的miR-148a-3p與賁門癌淋巴結(jié)轉(zhuǎn)移有著明顯的相關(guān)性,并預(yù)示著較差的總體生存期。
[Abstract]:Objective: to detect the expression profiles of long chain noncoding RNA (long non-coding) and microRNAs (microRNAs miRNAs) in gastric cardia carcinoma and paracancerous tissues by gene chip technique. To investigate the relationship between the differential expression of lnc RNA and miRNAs in gastric cardia carcinoma and paracancerous tissues, and the relationship between abnormal expression of miRNAs and prognosis. Materials and methods: a total of 52 surgically resected cardia carcinoma and paracancerous tissues were selected as study specimens, of which 15 samples were used to detect 37 cases of gastric cardia carcinoma by gene chip for late PCR verification. The Arraystar human ncRNA microarray V2.0 and miRCURYLNAmicro micro RNA microRNAs were used to detect the expression profiles of lncRNA and miRNA by Arraystar labeling and hybridization. The differentially expressed LNNs and miRNAs were obtained from cardiac carcinoma and paracancerous tissues. Bioinformatics prediction technique was used to analyze the correlation between partial lnc RNA and mi RNA in the expression profile, and the results were verified by qRT-PCR in 37 gastric cardia carcinomas and paracancerous tissues. The correlation between the expression of lncRNA and miRNA was analyzed statistically. The relationship between miRNA and clinicopathological features was analyzed by chi-square test, and the relationship between miRNA and survival time was studied by Kaplan-meier survival analysis. Results in 15 cases of gastric cardia carcinoma and adjacent tissues, there were more than 2 times of differentially expressed lncRNAs miRNAs and their mRNA expression profiles: a total of 2090 miRNAs and 152mRNAs were detected. According to the different expression multiple, standardized intensity value and annotated data of lncRNA, two lnc RNAs: H19 and MALAT1 were selected. By bioinformatics prediction, three miRNAs were found to be related to the expression of H19 and MALAT1 in cardiac carcinoma, including: has-mi R-148a-3pnhsa-miR-205-5p and hsa-miR-363-3p. The expression of H19 and has-miR-148a-3p were detected by qRT-PCR. The results showed that the expression of H19 in 37 patients with cardiac carcinoma and adjacent cancer was significantly lower than that in the control group (P0. 0072) mi R-148a-3p (P0.0001). The expression of miR-148a-3p was negatively correlated with the expression of H19 by nonparametric rank sum test (P0. 019). The analysis of clinicopathological features showed that the low expression of miRNA was significantly correlated with lymph node metastasis of cardiac carcinoma (P0. 039). Kaplan-meier survival analysis showed that the low expression of miR-148a-3p was a predictor of poor survival time of patients with gastric cardia carcinoma (P0. 022). Conclusion there is a significant difference in lnc and miRNA expression profiles between cardia carcinoma and paracancerous tissues. By means of bioinformatics prediction and qRT-PCR, the expression of lncRNA H19 and miR-148a-3p in gastric cardia carcinoma was found to be related to the expression of LncRNA H19 and miR-148a-3p. In addition, the low expression of miR-148a-3p was significantly correlated with lymph node metastasis of cardiac carcinoma, and predicted poor overall survival.
【學(xué)位授予單位】：河南科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R735.2

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