β2 毒素处理的 IPEC-J2 细胞中长非编码 RNA 和 mRNAs 的 N6-甲基腺苷甲基化分析。
N6-Methyladenosine Methylation Analysis of Long Noncoding RNAs and mRNAs in IPEC-J2 Cells Treated With beta2 Toxin.
机构信息
College of Animal Science and Technology, Gansu Agricultural University, Lanzhou, China.
College of Animal Science and Technology, Northwest A&F University, Xian, China.
出版信息
Front Immunol. 2021 Nov 22;12:769204. doi: 10.3389/fimmu.2021.769204. eCollection 2021.
BACKGROUND
The n6-methyladenosine (m6A) modification is present widely in mRNAs and long non-coding RNAs (lncRNAs), and is related to the occurrence and development of certain diseases. However, the role of m6A methylation in infectious diarrhea remains unclear.
METHODS
Here, we treated intestinal porcine jejunum epithelial cells (IPEC-J2 cells) with beta2 (CPB2) toxin to construct an model of infectious diarrhea, and then used methylated RNA immunoprecipitation sequencing (MeRIP-seq) and RNA sequencing (RNA-seq) to identify the methylation profiles of mRNAs and lncRNAs in IPEC-J2 cells.
RESULTS
We identified 6,413 peaks, representing 5,825 m6A-modified mRNAs and 433 modified lncRNAs, of which 4,356 m6A modified mRNAs and 221 m6A modified lncRNAs were significantly differential expressed between the control group and CPB2 group. The motif GGACU was enriched significantly in both the control group and the CPB2 group. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation analysis showed that the differentially methylated modified mRNAs were mainly enriched in Hippo signaling pathway and Wnt signaling pathway. In addition, the target genes of the differentially m6A modified lncRNAs were related to defense response to virus and immune response. For example, , and might regulate the defense response to virus, immune and inflammatory response to resist the harmful effects of viruses on cells.
CONCLUSION
In summary, this study established the m6A transcription profile of mRNAs and lncRNAs in IPEC-J2 cells treated by CPB2 toxin. Further analysis showed that m6A-modified RNAs were related to defense against viruses and immune response after CPB2 toxin treatment of the cells. Threem6A-modified lncRNAs, , and , were most likely to play a key role in CPB2 toxin-treated IPEC-J2 cells. The results provide a theoretical basis for further research on the role of m6A modification in piglet diarrhea.
背景
N6-甲基腺苷（m6A）修饰广泛存在于 mRNA 和长非编码 RNA（lncRNA）中，与某些疾病的发生和发展有关。然而，m6A 甲基化在感染性腹泻中的作用尚不清楚。
方法
在这里，我们用β2（CPB2）毒素处理猪肠上皮细胞（IPEC-J2 细胞），构建感染性腹泻模型，然后用 m6A 修饰 RNA 免疫沉淀测序（MeRIP-seq）和 RNA 测序（RNA-seq）来鉴定 IPEC-J2 细胞中 mRNA 和 lncRNA 的甲基化谱。
结果
我们鉴定出 6413 个峰，代表 5825 个 m6A 修饰的 mRNA 和 433 个修饰的 lncRNA，其中 4356 个 m6A 修饰的 mRNA 和 221 个 m6A 修饰的 lncRNA在对照组和 CPB2 组之间差异表达显著。在对照组和 CPB2 组中， motif GGACU 均显著富集。基因本体（GO）和京都基因与基因组百科全书（KEGG）注释分析表明，差异甲基化修饰的 mRNA 主要富集在 Hippo 信号通路和 Wnt 信号通路。此外，差异 m6A 修饰 lncRNA 的靶基因与抗病毒防御反应和免疫反应有关。例如，和可能通过调节抗病毒防御反应、免疫和炎症反应来抵抗病毒对细胞的有害影响。
结论
总之，本研究建立了 CPB2 毒素处理的 IPEC-J2 细胞中 mRNA 和 lncRNA 的 m6A 转录谱。进一步分析表明，m6A 修饰的 RNA 与 CPB2 毒素处理细胞后的抗病毒防御和免疫反应有关。三个 m6A 修饰的 lncRNA、、和可能在 CPB2 毒素处理的 IPEC-J2 细胞中发挥关键作用。研究结果为进一步研究 m6A 修饰在仔猪腹泻中的作用提供了理论依据。
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