piRNA在肝细胞癌组织中的差异表达分析

摘要

目的 探讨piRNA在肝细胞肝癌发生、发展中的作用. 方法 对肝癌患者的癌及癌旁组织、转移及非转移的肝癌组织进行小RNA二代测序,将测序数据过滤掉常见RNA序列后去重复;用piRNA预测器预测可能的新piRNA,并和下载的已知piRNA合并,筛选得到差异明显的NRNA.以miranda算法预测相应靶基因,并对其进行富集分析和功能预测,并选取piRNA在LM3和PLC细胞中行进一步功能(迁移能力)实验及分析.两组间均数比较采用独立样本t检验. 结果 测序共获得66 772个NRNA(已知149个),发现癌和癌旁组织表达差异明显的有241个piRNAs,转移和非转移肿瘤组织表达差异明显的1 634个piRNAs.对差异piRNA的靶基因进行GO和KEGG通路分析,发现主要和细胞黏附相关.而对选取的一个piRNA (piR1/97)进行功能实验,显示其促进肝癌细胞的迁移(LM3:t=8.829,P＜0.05;PLC.t=7.318,P＜0.05).结论 NRNA在肝细胞肝癌患者的癌及癌旁组织、转移及非转移患者的肝癌组织中表达量有差异,并可能参与肝细胞肝癌的转移过程,更多的功能分析需要进一步研究和实验确证.%Objective To investigates the role of piwi-interacting RNAs (piRNA) in the occurrence and development of hepatocellular carcinoma.Methods Second-generation small RNA sequencing was performed on cancer and paracancerous tissues,metastatic and non-metastatic liver cancer tissues of patients with liver cancer,and the sequencing data were filtered out for the common RNA sequences to be repeated.The piRNA predictor was used to forecast the possible new piRNA merged with the downloaded known piRNA to screen out distinction.A miRanda algorithm was used to predict the corresponding target genes and functional enrichment analysis.piRNA was selected for experimental functional (migration) analysis.An independent t-test was used to compare means between the two groups.Results 66 772 piRNAs (known 149) were obtained by sequencing.241 piRNAs were found in cancer and paracancerous tissues,and 1 634 piRNAs were found in metastatic and non-metastatic tumors.Analysis of the GO and KEGG pathways of the target genes of differential piRNAs revealed that they were mainly involved in cell adhesion.An experimental functional analysis was performed on the selected Pirna (PIR1/97),which showed that it promoted the migration of hepatoma cells (LM3:t =8.829,P ＜ 0.05;PLC:t =7.318,P ＜ 0.05).Conclusion The expression levels of piRNA in hepatocellular carcinoma patients with cancer and paracancerous tissues,metastasis and non-metastatic liver cancer tissues are different and it could be entailed in the metastasis process of hepatocellular carcinoma.Hence,experimental functional analysis is required for research and experimental confirmation.