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General Research Article

高通量测序揭示N6-甲基腺苷修饰的LncRNA作为肝纤维化小鼠的潜在生物标志物

卷 23, 期 5, 2023

发表于: 03 July, 2023

页: [371 - 390] 页: 20

弟呕挨: 10.2174/1566523223666230606151013

价格: $65

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摘要

背景：N6-甲基腺苷 (m6A) 是真核 RNA 中最常见的内部修饰。长非编码RNA（lncRNA）是一种具有多种细胞功能的新型非编码调节分子。两者均与肝纤维化（LF）的发生、发展密切相关。然而，m6A 甲基化 lncRNA 在 LF 进展中的作用仍然很大程度上未知。 方法：本研究采用HE和Masson染色观察肝脏病理变化，采用m6A修饰RNA免疫沉淀测序（m6A-seq）系统评价LF小鼠lncRNAs的m6A修饰水平，meRIP-qPCR和RT-qPCR用于检测目标lncRNA的m6A甲基化水平和RNA表达水平。 结果：肝纤维化组织中313个lncRNA中共检测到415个m6A峰。 LF中有98个显着不同的m6A峰，位于84个lncRNA上，其中45.2%的lncRNA长度在200-400 bp之间。同时，这些甲基化lncRNA的前3条染色体分别是7号、5号和1号染色体。RNA测序在LF中鉴定出154个差异表达的lncRNA。 m6A-seq和RNA-seq联合分析发现，m6A甲基化和RNA表达水平发生显着变化的lncRNA有3个：lncRNA H19、lncRNA Gm16023和lncRNA Gm17586。随后验证结果显示，lncRNA H19和lncRNA Gm17586的m6A甲基化水平显着升高，而lncRNA Gm16023的m6A甲基化水平显着降低，且3个lncRNA的RNA表达显着降低。通过创建lncRNA-miRNA-mRNA调控网络，揭示了lncRNA H19、lncRNA Gm16023和lncRNA Gm17586在LF中可能的调控关系。 结论：本研究揭示了LF小鼠lncRNA独特的m6A甲基化模式，提示lncRNA的m6A甲基化修饰与LF的发生、发展有关。

关键词: N6-甲基腺苷，lncRNA，调节网络，肝纤维化，高通量测序，meRIP-qPCR。

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