METTL3-Mediated LncRNA <i>EN_42575</i> m6A Modification Alleviates CPB2 Toxin-Induced Damage in IPEC-J2 Cells
Long non-coding RNAs (lncRNAs) modified by n6-methyladenosine (m6A) have been implicated in the development and progression of several diseases. However, the mechanism responsible for the role of m6A-modified lncRNAs in <i>Clostridium perfringens</i> type C piglet diarrhea has remained l...
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| Format: | Article |
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MDPI AG
2023-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/6/5725 |
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| author | Jiaojiao Yang Qiaoli Yang Xiaoyu Huang Zunqiang Yan Pengfei Wang Xiaoli Gao Jie Li Shuangbao Gun |
| author_facet | Jiaojiao Yang Qiaoli Yang Xiaoyu Huang Zunqiang Yan Pengfei Wang Xiaoli Gao Jie Li Shuangbao Gun |
| author_sort | Jiaojiao Yang |
| collection | DOAJ |
| description | Long non-coding RNAs (lncRNAs) modified by n6-methyladenosine (m6A) have been implicated in the development and progression of several diseases. However, the mechanism responsible for the role of m6A-modified lncRNAs in <i>Clostridium perfringens</i> type C piglet diarrhea has remained largely unknown. We previously developed an in vitro model of CPB2 toxin-induced piglet diarrhea in IPEC-J2 cells. In addition, we previously performed RNA immunoprecipitation sequencing (MeRIP-seq), which demonstrated lncRNA <i>EN_42575</i> as one of the most regulated m6A-modified lncRNAs in CPB2 toxin-exposed IPEC-J2 cells. In this study, we used MeRIP-qPCR, FISH, EdU, and RNA pull-down assays to determine the function of lncRNA <i>EN_42575</i> in CPB2 toxin-exposed IPEC-J2 cells. LncRNA <i>EN_42575</i> was significantly downregulated at different time points in CPB2 toxin-treated cells. Functionally, lncRNA <i>EN_42575</i> overexpression reduced cytotoxicity, promoted cell proliferation, and inhibited apoptosis and oxidative damage, whereas the knockdown of lncRNA <i>EN_42575</i> reversed these results. Furthermore, the dual-luciferase analysis revealed that METTL3 regulated lncRNA <i>EN_42575</i> expression in an m6A-dependent manner. In conclusion, METTL3-mediated lncRNA <i>EN_42575</i> exerted a regulatory effect on IPEC-J2 cells exposed to CPB2 toxins. These findings offer novel perspectives to further investigate the function of m6A-modified lncRNAs in piglet diarrhea. |
| first_indexed | 2024-03-11T06:25:03Z |
| format | Article |
| id | doaj.art-677e8d8ff89545f4beac27c00f1b9d8b |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T06:25:03Z |
| publishDate | 2023-03-01 |
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| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-677e8d8ff89545f4beac27c00f1b9d8b2023-11-17T11:38:08ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-03-01246572510.3390/ijms24065725METTL3-Mediated LncRNA <i>EN_42575</i> m6A Modification Alleviates CPB2 Toxin-Induced Damage in IPEC-J2 CellsJiaojiao Yang0Qiaoli Yang1Xiaoyu Huang2Zunqiang Yan3Pengfei Wang4Xiaoli Gao5Jie Li6Shuangbao Gun7College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaCollege of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaCollege of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaCollege of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaCollege of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaCollege of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaCollege of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaCollege of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, ChinaLong non-coding RNAs (lncRNAs) modified by n6-methyladenosine (m6A) have been implicated in the development and progression of several diseases. However, the mechanism responsible for the role of m6A-modified lncRNAs in <i>Clostridium perfringens</i> type C piglet diarrhea has remained largely unknown. We previously developed an in vitro model of CPB2 toxin-induced piglet diarrhea in IPEC-J2 cells. In addition, we previously performed RNA immunoprecipitation sequencing (MeRIP-seq), which demonstrated lncRNA <i>EN_42575</i> as one of the most regulated m6A-modified lncRNAs in CPB2 toxin-exposed IPEC-J2 cells. In this study, we used MeRIP-qPCR, FISH, EdU, and RNA pull-down assays to determine the function of lncRNA <i>EN_42575</i> in CPB2 toxin-exposed IPEC-J2 cells. LncRNA <i>EN_42575</i> was significantly downregulated at different time points in CPB2 toxin-treated cells. Functionally, lncRNA <i>EN_42575</i> overexpression reduced cytotoxicity, promoted cell proliferation, and inhibited apoptosis and oxidative damage, whereas the knockdown of lncRNA <i>EN_42575</i> reversed these results. Furthermore, the dual-luciferase analysis revealed that METTL3 regulated lncRNA <i>EN_42575</i> expression in an m6A-dependent manner. In conclusion, METTL3-mediated lncRNA <i>EN_42575</i> exerted a regulatory effect on IPEC-J2 cells exposed to CPB2 toxins. These findings offer novel perspectives to further investigate the function of m6A-modified lncRNAs in piglet diarrhea.https://www.mdpi.com/1422-0067/24/6/5725m6AlncRNA <i>EN_42575</i>CPB2 toxinIPEC-J2 cellMETTL3 |
| spellingShingle | Jiaojiao Yang Qiaoli Yang Xiaoyu Huang Zunqiang Yan Pengfei Wang Xiaoli Gao Jie Li Shuangbao Gun METTL3-Mediated LncRNA <i>EN_42575</i> m6A Modification Alleviates CPB2 Toxin-Induced Damage in IPEC-J2 Cells International Journal of Molecular Sciences m6A lncRNA <i>EN_42575</i> CPB2 toxin IPEC-J2 cell METTL3 |
| title | METTL3-Mediated LncRNA <i>EN_42575</i> m6A Modification Alleviates CPB2 Toxin-Induced Damage in IPEC-J2 Cells |
| title_full | METTL3-Mediated LncRNA <i>EN_42575</i> m6A Modification Alleviates CPB2 Toxin-Induced Damage in IPEC-J2 Cells |
| title_fullStr | METTL3-Mediated LncRNA <i>EN_42575</i> m6A Modification Alleviates CPB2 Toxin-Induced Damage in IPEC-J2 Cells |
| title_full_unstemmed | METTL3-Mediated LncRNA <i>EN_42575</i> m6A Modification Alleviates CPB2 Toxin-Induced Damage in IPEC-J2 Cells |
| title_short | METTL3-Mediated LncRNA <i>EN_42575</i> m6A Modification Alleviates CPB2 Toxin-Induced Damage in IPEC-J2 Cells |
| title_sort | mettl3 mediated lncrna i en 42575 i m6a modification alleviates cpb2 toxin induced damage in ipec j2 cells |
| topic | m6A lncRNA <i>EN_42575</i> CPB2 toxin IPEC-J2 cell METTL3 |
| url | https://www.mdpi.com/1422-0067/24/6/5725 |
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