| 摘要: |
| ［摘要］ 目的 探究长链非编码RNA（lncRNA）肝癌高表达转录本（HULC）对肝癌细胞微小RNA（miRNA）差异表达谱及竞争内源性RNA（ceRNA）调控网络的影响。方法 选取人肝癌细胞系BEL-7402，瞬时转染HULC siRNA以降低lncRNA HULC的表达。通过高通量测序选取表达具有差异的miRNA，并通过实时荧光定量PCR进行验证。基于参考基因组，预测新miRNA。通过基因富集分析及ceRNA调控网络，研究这些基因的潜在功能。结果 经敲低lncRNA HULC表达后，通过高通量测序挑选出了9个表达差异显著的miRNA。5个miRNA表达下调，4个miRNA表达上调。通过miRDeep2打分，挑选出了15个分值较高的新miRNA。基因富集分析结果显示，表达下调的miRNA的靶基因的分子功能与丝氨酸tRNA连接酶活性、蛋白激酶活性、阳离子通道活性、钙转运ATP酶活性等有关；表达上调的miRNA的靶基因与硒代半胱氨酸裂解酶活性、过氧化物酶体机制靶向信号1结合、过氧化物酶体靶向序列结合、蛋白质N-末端结合、肿瘤坏死因子受体超家族结合等有关。miRNA-靶基因互作网络、显著富集功能-功能互作网络揭示靶基因与miRNA的关联性以及功能间的相关性。结论 在肝癌细胞系BEL-7402中lncRNA HULC具有调节多个表达差异的miRNA的作用，通过ceRNA调控网络模式，影响肝癌的发生、发展。 |
| 关键词: 长链非编码RNA 肝癌高表达转录本 肝细胞癌 高通量测序 微小RNA |
| DOI：10.3969/j.issn.1674-3806.2022.05.06 |
| 分类号:R 735.7 |
| 基金项目:河南省重点研发与推广专项（科技攻关）项目（编号：202102310126）；河南省医学科技攻关计划联合共建项目（编号：LHGJ20191342） |
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| Effects of lncRNA HULC on miRNA differential expression profile and ceRNA regulatory network in hepatocarcinoma cells |
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GUAN Lu-lu, HAN Song-feng, LI Shi-peng, et al.
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Medical Research Center, Basic Medical College, Henan University of Science and Technology, Luoyang 471000, China
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| Abstract: |
| ［Abstract］ Objective To explore the effects of long non-coding ribonucleic acid(lncRNA) highly up-regulated in liver cancer(HULC) on micro ribonucleic acid(miRNA) differential expression profile and competitive endogenous ribonucleic acid(ceRNA) regulatory network of liver cancer cells. Methods Human liver cancer cell line BEL-7402 was selected, and HULC siRNA was transfected instantaneously into human liver cancer cell line BEL-7402 to reduce the expression of lncRNA HULC. The different expressions of miRNA were selected by using high-throughput sequencing and were verified by real-time quantitative reverse transcription polymerase chain reaction(qRT-PCR). The new miRNA was predicted based on reference genomes. Gene enrichment analysis and ceRNA regulatory network were used to study the potential function of these genes. Results After the expression of lncRNA HULC was knocked down, nine significantly differentially expressed miRNAs were selected by high-throughput sequencing. The expressions of 5 miRNAs were down-regulated and those of 4 miRNAs were up-regulated. Fifteen new miRNAs with higher scores were selected by using miRDeep2 scoring. The results of gene enrichment analysis showed that the molecular functions of the down-regulated miRNA target genes were related to the activities of serine tRNA ligase, protein kinase, cationic channel and calcium transporter ATPase. The target genes of the up-regulated miRNAs were related to selenocysteine lyase activity, peroxisomal mechanism targeting signal-1 binding, peroxisomal targeting sequence binding, protein N-tail binding, tumor necrosis factor receptor superfamily binding, etc. The miRNA-target gene interaction network and the significantly enriched function-function interaction network revealed the association of target genes and miRNA, and the correlation between functions. Conclusion In liver cancer cell line BEL-7402, lncRNA HULC has the role of regulating multiple miRNAs with different expressions and has effects on the occurrence and development of liver cancer through the regulation network mode of ceRNA. |
| Key words: Long non-coding ribonucleic acid(lncRNA) Highly up-regulated in liver cancer(HULC) Hepatocellular carcinoma(HCC) High-throughput sequencing Micro ribonucleic acid(miRNA) |
