^(125)I放射性粒子诱导肝癌细胞长链非编码RNA表达谱变化研究

Study on the changes in the long non-coding RNA expression profile of hepatocarcinoma cells induced by^(125)I radioactive particles

目的探讨^(125)I放射性粒子辐射诱导人肝癌细胞(Hep3B)长链非编码RNA(lncRNA)表达谱的变化。方法采用Illumina PE150平台对^(125)I放射性粒子辐照和未经辐照的Hep3B细胞进行lncRNA测序,对原始数据进行lncRNA表达量、差异lncRNA、GO功能富集等生物信息分析;选取表达差异显著的lncRNAs,设计引物进行实时荧光定量聚合酶链式反应(qRT-PCR)验证测序结果。结果实验中得到9206个差异lncRNAs转录本,其中上调4556个,下调4650个;6个表达差异较大基因的qRT-PCR验证结果与测序结果一致,显示测序结果可靠;生物信息分析发现差异lncRNAs转录本的靶基因功能主要集中在自噬体形成、DNA损伤等方面,主要参与细胞增殖的调控。结论^(125)I放射性粒子诱导肝癌Hep3B细胞lncRNA表达谱发生变化,为进一步探索电离辐射诱导肿瘤细胞死亡分子机制、寻找放疗生物标志物和干预靶点提供了新思路。

Objective To explore the changes in the long non-coding RNA(lncRNA)expression profile of human hepatocellular carcinoma cells(Hep3B)induced by^(125)I radioactive particles.Methods The lncRNA sequencing of Hep3B cells irradiated or unirradiated by^(125)I radioactive particles was performed in the Illumina PE150 platform.The original sequencing data were analysed by the lncRNA expression,differential lncRNA expression,GO function enrichment carried out,and other bioinformatics.The lncRNAs with significant differences in expression were selected,and primers targeting of lncRNAs were designed to verify the sequencing results by Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).Results A total of 9206 differential lncRNA transcripts were obtained from the sequencing experiment,of which 4556 were up-regulated and 4650 down-regulated.The qRT-PCR verification results of 6 genes with remarked expression differences were consistent with the sequencing results,indicating that the sequencing results were reliable.Bioinformatics analyses showed that the target genes of differential lncRNA transcripts focus on autophagy formation,DNA damage,which were mainly involved in the regulation of cell proliferation.Conclusion^(125)I radioactive particles induce changes in the lncRNA expression profile of liver cancer Hep3B cells,which provides new ideas for further exploring the molecular mechanism of ionizing radiation-induced tumor cell death,finding radiotherapy biomarkers and intervention targets.