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中国应用生理学杂志 ›› 2021, Vol. 37 ›› Issue (4): 343-348.doi: 10.12047/j.cjap.6076.2021.038

• 研究论文 • 上一篇    下一篇

低氧mtDNA3010A/G基因型变异诱导的lncRNA-mRNA共表达网络变化

陈郁, 龚亮, 罗勇军   

  1. 陆军军医大学陆军卫勤训练基地军事医学地理学教研室, 重庆 400038
  • 出版日期:2021-07-28 发布日期:2021-08-09
  • 通讯作者: Tel: 023-68771868; E-mail: ajun-333333@163.com
  • 基金资助:
    *国家自然科学基金面上项目(41877518);陆军军医大学青年培育项目(2016XPY09);陆军军医大学科技创新能力提升专项项目(2019XYY09);青海省基础研究计划项目(2018-ZJ-705)

Changes of lncRNA-mRNA co-expression network induced by mtDNA3010A/G genotype mutation in hypoxia

CHEN Yu, GONG Liang, LUO Yong-jun   

  1. Department of Military Medical Geography, Army Health Service Training Base, Army Medical University, Chongqing 40038, China
  • Online:2021-07-28 Published:2021-08-09

摘要: 目的: 分析mtDNA3010A/G变异在急性缺氧条件下的长链非编码RNA(lncRNA)和信使RNA(mRNA)的共表达网络变化,探讨关键lncRNA和mRNA在低氧诱导的基因表达调控中的作用。方法: 筛选线粒体DNA(mtDNA)3010-5178-10400的基因型组合A-C-C和G-C-C,以骨肉瘤细胞经溴化乙锭处理后形成的无线粒体细胞(ρ0206细胞)为供体,构建mtDNA3010A和mtDNA3010G基因型融合细胞。经1%O2处理24 h后,采用lncRNA-mRNA共表达芯片检测两种融合细胞的差异表达lncRNA和mRNA,荧光定量聚合酶链式法验证差异显著的mRNA,运用生物信息学方法构建lncRNA-mRNA共表达网络,预测差异lncRNA的靶基因,并对差异显著的mRNA和预测靶基因进行基因本体(GO)和京都基因与基因组大百科全书(KEGG)预测分析。结果: 经1%O2处理24 h后,与mtDNA3010G融合细胞相比,mtDNA3010A融合细胞表达上调的lncRNA有688个,超过2倍的有21个,表达下调的lncRNA有1098个,超过2倍的有4个;表达上调的mRNA有1151个,超过2倍的有14个,表达下调的mRNA有539个,超过2倍的有3个。结论: mtDNA3010A/G基因型变异在缺氧条件下能够影响lncRNA-mRNA调控网络的变化,差异表达的lncRNA和mRNA可能在低氧诱导的基因表达调控网络中发挥重要作用,有望成为从线粒体角度调控低氧反应的靶点。

关键词: 线粒体DNA, 变异, 长链非编码RNA, 低氧反应, 共表达网络

Abstract: Objective: To analyze the long non-coding RNA (lncRNA) and messenger RNA (mRNA) co-expression network changes induced by mtDNA3010A/G mutation in acute hypoxia, and to investigate the role of key lncRNA and mRNA in the regulation of gene expression induced by hypoxia. Methods: The genotype combinations A-C-C and G-C-C of mitochondrial DNA 3010-5178-10400 were screened, and genotypes of mtDNA3010A and mtDNA3010G fusion cells were constructed by using osteosarcoma cell treated by ethidium bromide without mitochondrion (ρ0206 cell) as donors. After treatment with 1% O2 24 h, the lncRNA - mRNA expression chip was applied to detect the differently expressed lncRNA and mRNA in two kinds of fusing cells, and fluorescence quantitative polymerase chain method was used to verify differently expressed mRNA. Bioinformatics methods were applied to build co-expression network of lncRNA-mRNA, predict target genes of differently expressed lncRNA, and the functions of differently expressed mRNA and target genes predicted by lncRNA were also analyzed based on gene ontology (GO) and the Kyoto encyclopedia of genes and genomes (KEGG) forecast analysis. Results: After treatment with 1% O2 for 24 h, compared with mtDNA3010G fusion cells: 688 lncRNAs were up-regulated, 21 were more than 2 times; 1098 were down-regulated, and 4 were more than 2 times. There were 1151 mRNA expressions up-regulated, 14 were more than 2 times, 539 mRNA expressions were down-regulated, and 3 were more than 2 times. Conclusion: MtDNA3010A/G genotype mutation under hypoxia is able to affect the lncRNA-mRNA regulatory network, and the differentially expressed lncRNA and mRNA may play an important role in regulation network of gene expression induced by hypoxia, which is expected to be a target for the regulation of hypoxia reaction from the perspective of mitochondria.

Key words: mitochondrial DNA, variation, long non-coding RNA, hypoxic response, co-expression network

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