不同类型肝内胆管细胞癌长链非编码RNA差异表达分析及意义

发布时间：2018-04-23 13:09

本文选题：肝内胆管结石 + 肝内胆管细胞癌　；参考：《宁波大学》2017年硕士论文

【摘要】：目的:肝内胆管细胞癌是一种死亡率较高的恶性肿瘤,其发病与肝内胆管结石密切相关。越来越多的证据表明,长链非编码RNA参与多种肿瘤的发生发展过程。然而它在肝内胆管细胞癌(包括结石相关性及非结石相关性肝内胆管细胞癌)中的作用目前仍不清楚。因此,本研究旨在探究不同类型肝内胆管细胞癌的分子机制差异,并探索结石相关性肝内胆管细胞癌相关的长链非编码RNA。方法:通过基因芯片检测结石相关性及非结石相关性肝内胆管癌的lnc RNA及m RNA表达情况,筛选肿瘤相关的差异基因。利用GO基因富集分析和KEGG通路分析进一步分析差异表达的m RNA在两种类型肿瘤中的功能富集及肿瘤相关信号通路。实时荧光定量PCR用于检测lnc RNA-AFAP1-AS1在两种类型肝内胆管细胞癌的表达情况并分析其表达水平与临床病理特征的相关性。此外,通过m RNA-lnc RNA共表达网络构建及共表达基因功能富集分析预测lnc RNA-AFAP1-AS1功能。结果:基因芯片分析结果显示两种不同类型的肝内胆管细胞癌的lnc RNA和m RNA的表达模式存在差异。GO富集分析和KEGG通路分析结果亦提示结石相关性肝内胆管细胞癌的分子机制不同于非结石相关性肝内胆管细胞癌。通过比较筛选出两类肝内胆管细胞癌中表达不同的差异基因,这些可能反映了它们不同的发病机制。实时荧光定量PCR结果表明,AFAP1-AS1在结石相关性肝内胆管细胞癌的肿瘤组织中表达水平高于癌旁组织,而在非结石相关性肝内胆管细胞癌中差异无统计学意义。高水平的AFAP1-AS1与肿瘤大小相关。共表达基因富集分析预测其功能主要涉及细胞运动迁移、上皮细胞分化及调节免疫系统过程。结论:本研究首次描述结石相关性肝内胆管细胞癌中lnc RNA及m RNA的表达情况,为进一步寻找生物标志物提供有价值的信息。同时证实AFAP1-AS1在结石相关性肝内胆管细胞癌中的表达水平上调,其可能在结石致癌过程中发挥重要作用。
[Abstract]:Objective: intrahepatic cholangiocarcinoma is a malignant tumor with high mortality. There is growing evidence that long-chain non-coding RNA is involved in the development of multiple tumors. However, its role in intrahepatic cholangiocarcinoma (including stone-associated and non-calculous-associated intrahepatic cholangiocarcinoma) remains unclear. Therefore, the purpose of this study was to explore the molecular mechanisms of different types of intrahepatic cholangiocarcinoma and to explore the long chain noncoding RNA associated with lithiasis associated intrahepatic cholangiocarcinoma. Methods: the expression of lnc RNA and m RNA in lithiasis related and non-stone associated intrahepatic cholangiocarcinoma were detected by gene microarray, and the differentially expressed genes were screened. Go gene enrichment analysis and KEGG pathway analysis were used to further analyze the functional enrichment and tumor-related signaling pathway of differentially expressed m RNA in two types of tumors. Real-time fluorescence quantitative PCR was used to detect the expression of lnc RNA-AFAP1-AS1 in two types of intrahepatic cholangiocarcinoma and to analyze the correlation between the expression level and clinicopathological features. In addition, the m RNA-lnc RNA coexpression network was constructed and the function of lnc RNA-AFAP1-AS1 was predicted by functional enrichment analysis of coexpression genes. Results: the results of gene chip analysis showed that the expression patterns of lnc RNA and m RNA in two different types of intrahepatic cholangiocarcinoma were different. Go enrichment analysis and KEGG pathway analysis also suggested that lithiasis associated intrahepatic cholangiocarcinoma. The molecular mechanism is different from that of non-stone-associated intrahepatic cholangiocarcinoma. The differentially expressed genes in two types of intrahepatic cholangiocarcinoma were screened, which may reflect their different pathogenesis. The results of real-time fluorescence quantitative PCR showed that AFAP1-AS1 expression was higher in lithium-associated intrahepatic cholangiocarcinoma than that in paracancerous tissue, but there was no significant difference in non-stone associated intrahepatic cholangiocarcinoma. High levels of AFAP1-AS1 are associated with tumor size. The function of co-expression gene enrichment analysis is mainly related to cell migration, epithelial cell differentiation and regulation of immune system. Conclusion: this study first describes the expression of lnc RNA and m RNA in lithiasis associated intrahepatic cholangiocarcinoma and provides valuable information for further searching for biomarkers. It is also confirmed that the expression of AFAP1-AS1 in lithiasis associated intrahepatic cholangiocarcinoma is up-regulated, which may play an important role in the carcinogenesis of calculi.
【学位授予单位】：宁波大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.8

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