特异长链非编码RNA在肝细胞癌中的表达和验证  

Expression and verification of specific long non-coding RNA in hepatocellular carcinoma

作　　者：厉新妍[1] 方亮[1] 黄淑琳[2] 

机构地区：[1]广州市第一人民医院,广东广州510180 [2]香港大学理学院

出　　处：《广州医科大学学报》2016年第5期1-8,共8页Academic Journal of Guangzhou Medical University

基　　金：广东省科技计划项目(2012B031800010)

摘　　要：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 the differentially expressed of long non-coding RNA（ lncRNA） in hepatocellular carcinoma（ HCC） tissues and matched adjacent normal tissues,and to investigate the specific lncRNA that may be involved in the occurrence and development of HCC. Methods： lncRNA and mRNA were determined in 5 cases of HCC and the matched adjacent normal tissues using lncRNA chip technique,and the differentially expressed lncRNA and mRNA were screened out,and classified by bioinformatics techniques of hierarchical clustering analysis,GO analysis and pathway analysis. Finally,10 lncRNAs were selected for quantitative validation by RT-PCR. Results： There were significant differences in the expression levels of 1 359 lncRNAs,with a fold change 〉2-fold. A total of 629 lncRNAs were significantly up-regulated,accounting for46.2%,of which 125 were more than 5-fold; 730 were down-regulated,accounting for 53.7%,110 of which were more than 5-fold. Cluster analysis：（1） there were 11 706 lncRNAs between genes,among which 352 lncRNAs were〈300 kb from the known coding genes.（2） There were 1 802 enhancer-type lncRNAs,of which there were 42 lncRNAs with〈300 kb distance from the known coding genes.（3） There were 101 HOX gene clusters. GO analysis divided the mRNA into molecular function,biological process and cellular component. Pathway analysis： There were 61 pathways in the differentially expressed mRNA genes involved regulation of cell life activity. Up-regulated mRNA was involved in 12 pathways,and down-regulated mRNA was involved in 49 pathways. The expression levels of the lncRNAs involved in the cell cycle and chemical carcinogenesis were significantly changed. RTPCR： The expression trends of 8 lncRNAs were consistent with the gene chip. There were 6 significant differences between the HCC group and the matched adjacent normal group（ P〈0.05）. Conclusion： The expression level of lncRNA in HCC is significantly changed compared with that in adjacent normal tissues,which may be relate

关 键 词：肝细胞癌 长链非编码RNA 长链非编码RNA芯片 

分 类 号：R735.7[医药卫生—肿瘤]