论文部分内容阅读
目的:分析肝细胞癌组织与配对癌旁组织中表达差异的长链非编码RNA(lncRNA),探讨可能与肝细胞癌发生发展有关的特异lncRNA。方法:采用lncRNA芯片技术,检测5例肝细胞癌和其配对癌旁正常组织中的lncRNA和mRNA,筛选出差异表达的lncRNA和mRNA。运用分层聚类分析,GO分析和pathway分析等生物信息学技术对其进行分类。最后从中选择10个lncRNA用RT-PCR进行定量验证。结果:有1 359个lncRNA表达水平出现显著性差异,差异倍数>2倍。629个显著上调,占46.2%,其中125个表达5倍以上;730个显著下调,占53.7%,其中110个表达5倍以上。聚类分析结果:(1)基因间lncRNA有11 706条,其中与已知编码基因相距<300 kb的lncRNA共有352条。(2)增强子型lncRNA共有1 802条,其中与已知编码基因相距<300 kb的lncRNA共有42条。(3)HOX基因簇共有101条。GO分析将mRNA分成分子功能、生物过程和细胞组分3类。Pathway分析结果:差异表达的mRNA基因所参与的细胞生命活动调控通路共有61条,上调表达的mRNA参与12条通路,下调表达的mRNA参与49条通路。参与细胞周期和化学致癌作用的表达水平明显改变。RT-PCR结果:8条lncRNA表达趋势与基因芯片结果一致。对癌组和癌旁组进行检验,有6条有显著性差异(P<0.05)。结论:与癌旁组织比较,lncRNA肝细胞癌组织中的表达水平明显改变,可能与肝细胞癌的发生发展有关。
OBJECTIVE: To analyze long-chain non-coding RNA (lncRNA) that is differentially expressed in hepatocellular carcinoma tissues and paracancerous tissues, and to explore specific lncRNAs that may be involved in the development of hepatocellular carcinoma. Methods: lncRNA microarray was used to detect lncRNA and mRNA in 5 cases of hepatocellular carcinoma (HCC) and its adjacent normal tissues, and the differentially expressed lncRNA and mRNA were screened out. Hierarchical cluster analysis, GO analysis and pathway analysis and other bioinformatics techniques to classify them. Finally, ten lncRNAs were selected for quantitative verification by RT-PCR. Results: There were 1 359 lncRNA expression levels were significantly different, the difference multiple> 2 times. 629 were significantly up-regulated, accounting for 46.2%, of which 125 were expressed more than 5 times; 730 were significantly down-regulated, accounting for 53.7% of which 110 were expressed more than 5 times. Results of cluster analysis: (1) There were 11 706 lncRNAs among which there were 352 lncRNAs <300 kb away from the known coding genes. (2) There were 1 802 lines of enhancer lncRNA, of which there were 42 lncRNAs <300 kb away from the known coding genes. (3) There are 101 HOX gene clusters in total. GO analysis divides mRNA into molecular functions, biological processes and cellular components. Pathway analysis results showed that there were 61 regulatory pathways involved in cell life activities of differentially expressed mRNAs, up-regulated mRNAs involved in 12 pathways and down-regulated mRNAs involved in 49 pathways. The levels of expression involved in cell cycle and chemical carcinogenesis are significantly altered. RT-PCR results: The ten lncRNA expression trends consistent with the results of the gene chip. There were 6 significant differences between the cancer group and the adjacent non-cancerous group (P <0.05). Conclusion: The expression level of lncRNA in hepatocellular carcinoma significantly changes compared with the adjacent tissues, which may be related to the occurrence and development of hepatocellular carcinoma.