贲门癌中长链非编码RNA和微小RNA表达谱及相关性分析

发布时间：2018-07-10 11:01

  本文选题：贲门癌 + 长链非编码RNA　； 参考：《河南科技大学》2017年硕士论文

【摘要】：目的:利用基因芯片技术检测贲门癌与癌旁组织中长链非编码RNA(long non-coding RNA,lnc RNA)和微小RNA(microRNA,miRNA)表达谱;探讨贲门癌与癌旁组织中差异性表达lnc RNA和miRNA之间的相关性以及异常表达mi RNA与预后的关系。材料与方法:共选取52对经手术切除的贲门癌与癌旁组织作为研究标本,其中15例标本用于基因芯片检测,37例标本进行后期PCR验证。利用Arraystar Human LncRNA microarray V2.0和miRCURY LNA?micro RNA Array,7th gen-hsa,mmurno两种芯片来检测lncRNA和miRNA的表达谱,再通过RNA标记与杂交、数据分析后得出贲门癌与癌旁组织中差异性表达的lncRNA和miRNA的表达谱。应用生物信息预测技术来分析表达谱中部分lnc RNA和mi RNA的相关性,并运用qRT-PCR技术在37对贲门癌与癌旁组织中验证结果。运用统计学分析lncRNA和miRNA两者之间的表达相关性。利用卡方检验来分析mi RNA与临床病理特征之间的关系;运用Kaplan-meier生存分析来研究miRNA与生存期的关系。结果:15例贲门癌与癌旁组织中2倍以上差异性表达的lncRNA、miRNA及mRNA的表达谱:lncRNA共计2090个;miRNA共计152。根据lncRNA表达谱中差异性表达倍数、标准化强度值及lncRNA数据的注释,选出两条lnc RNA:H19和MALAT1。通过生物信息预测方法,从贲门癌miRNA表达谱找到与H19和MALAT1存在表达相关性的miRNA共3条,包括:has-mi R-148a-3p、hsa-miR-205-5p及hsa-miR-363-3p。利用qRT-PCR技术验证一对预测的RNA:H19和has-miR-148a-3p的表达情况,结果显示在37例贲门癌与癌旁组H19呈明显的过表达(P=0.0072),mi R-148a-3p呈现明显的低表达(P0.0001)。利用非参数秩和检验发现H19与miR-148a-3p的表达呈现明显的负相关性(P=0.019)。临床病理特征分析提示低表达的miRNA与贲门癌淋巴结转移明显相关(P=0.039)。Kaplan-meier生存分析发现低表达的miR-148a-3p预示着较差的贲门癌患者生存期(P=0.022)。结论:15对贲门癌与癌旁组织的lnc RNA及miRNA表达谱存在显著差异。通过生物信息预测和qRT-PCR技术,找到了贲门癌中lncRNA H19和miR-148a-3p存在表达相关性。此外,低表达的miR-148a-3p与贲门癌淋巴结转移有着明显的相关性,并预示着较差的总体生存期。
[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.
【学位授予单位】：河南科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.2

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