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目的分析肝细胞癌与正常肝组织中差异表达的长链非编码RNA(lncRNA),发现可能与肝细胞癌发生发展相关的特异lncRNA。方法采用lncRNA表达谱芯片技术检测5例肝细胞癌和其配对癌旁正常组织中的lncRNA,筛选出差异表达的lncRNA。运用分层聚类分析,对所有的lncRNA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1359个lncRNA表达水平出现显著性差异,差异倍数〉2倍。其中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(1)基因间lncRNA有11706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1802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。RT-PCR结果有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条差异有统计学意义(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 differentially expressed long-chain non-coding RNA (lncRNA) in hepatocellular carcinoma (HCC) and normal liver tissues and to identify specific lncRNAs that may be associated with the development of hepatocellular carcinoma. Methods lncRNA expression profiling was used to detect lncRNA in 5 cases of hepatocellular carcinoma and its adjacent normal tissues, and the differentially expressed lncRNA was screened out. Using hierarchical clustering analysis, all lncRNAs were classified. Finally, ten lncRNAs were selected for quantitative verification by RT-PCR. Results There were 1359 lncRNA expression levels were significantly different, the difference multiple> 2 times. Among them, 629 were significantly up-regulated, accounting for 46.2%, of which 125 were more than 5-fold; 730 were significantly down-regulated, accounting for 53.7% of which 110 were more than 5-fold. Cluster analysis (1) There were 11,706 lncRNAs among which there were 352 lncRNAs <300 kb away from the known coding genes. (2) A total of 1802 lncRNAs were found in enhancer type, among which there were 42 lncRNAs that were <300 kb away from the known coding genes. (3) There are 101 HOX gene clusters in total. RT-PCR results showed that 8 lncRNA expression trends consistent with the results of the gene chip. There were 6 significant differences between the cancer group and the paracancer group (P <0.05). Conclusion Compared with the adjacent non-cancerous tissues, the expression level of lncRNA hepatocellular carcinoma significantly changes, which may be related to the occurrence and development of hepatocellular carcinoma.