METTL3-driven m6A modification of lncRNA FAM230B suppresses ferroptosis by modulating miR-27a-5p/BTF3 axis in gastric cancer

IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Yejia Cui , Meicen Pu , Yanting Gong , Runchao Li , Xiaokang Wang , Jinjun Ye , Haohai Huang , Dan Liao , Yufeng Yang , Aiping Yin , Jiale Li , Yuling Deng , Zhen Tian , Rong Pu
{"title":"METTL3-driven m6A modification of lncRNA FAM230B suppresses ferroptosis by modulating miR-27a-5p/BTF3 axis in gastric cancer","authors":"Yejia Cui ,&nbsp;Meicen Pu ,&nbsp;Yanting Gong ,&nbsp;Runchao Li ,&nbsp;Xiaokang Wang ,&nbsp;Jinjun Ye ,&nbsp;Haohai Huang ,&nbsp;Dan Liao ,&nbsp;Yufeng Yang ,&nbsp;Aiping Yin ,&nbsp;Jiale Li ,&nbsp;Yuling Deng ,&nbsp;Zhen Tian ,&nbsp;Rong Pu","doi":"10.1016/j.bbagen.2024.130714","DOIUrl":null,"url":null,"abstract":"<div><div>Our previous research revealed the apoptosis-inhibiting effect of lncRNA FAM230B in gastric cancer (GC). While its role on ferroptosis of GC remain unexplored. In this study, the m6A level and RNA stability regulation of METTL3 on FAM230B was detected by m6A quantification, stability assays, MeRIP, and their interaction was confirmed by RIP, and RNA pull-down assays. The level of ferroptosis was detected by flow cytometry, MDA and GSH level assessments, and electron microscopy. Gene expression was detected by quantitative real-time PCR, western blot, and immunofluorescence. The miR-27a-5p and BTF3 interaction was predicted with TargetScan and confirmed by dual-luciferase assay. Here, elevated levels of METTL3 and FAM230B were observed in GC tissues and cell lines. METTL3 was confirmed to bind with FAM230B RNA. Furthermore, silencing METTL3 reduced FAM230B m6A levels and stability, leading to decreased FAM230B and increased miR-27a-5p expressions. FAM230B knockdown favored ferroptosis and increased BTF3 expression, while its overexpression mitigated erastin-induced ferroptosis in GC cells. Additionally, BTF3 overexpression was found to negate miR-27a-5p's ferroptosis-promoting effects in GC cells. Collectively, our study demonstrates that the m6A modification of FAM230B by METTL3 plays a crucial role in promoting GC progression by reducing ferroptosis, through the modulation of the miR-27a-5p/BTF3 axis.</div></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":null,"pages":null},"PeriodicalIF":2.8000,"publicationDate":"2024-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biochimica et biophysica acta. General subjects","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0304416524001570","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

Our previous research revealed the apoptosis-inhibiting effect of lncRNA FAM230B in gastric cancer (GC). While its role on ferroptosis of GC remain unexplored. In this study, the m6A level and RNA stability regulation of METTL3 on FAM230B was detected by m6A quantification, stability assays, MeRIP, and their interaction was confirmed by RIP, and RNA pull-down assays. The level of ferroptosis was detected by flow cytometry, MDA and GSH level assessments, and electron microscopy. Gene expression was detected by quantitative real-time PCR, western blot, and immunofluorescence. The miR-27a-5p and BTF3 interaction was predicted with TargetScan and confirmed by dual-luciferase assay. Here, elevated levels of METTL3 and FAM230B were observed in GC tissues and cell lines. METTL3 was confirmed to bind with FAM230B RNA. Furthermore, silencing METTL3 reduced FAM230B m6A levels and stability, leading to decreased FAM230B and increased miR-27a-5p expressions. FAM230B knockdown favored ferroptosis and increased BTF3 expression, while its overexpression mitigated erastin-induced ferroptosis in GC cells. Additionally, BTF3 overexpression was found to negate miR-27a-5p's ferroptosis-promoting effects in GC cells. Collectively, our study demonstrates that the m6A modification of FAM230B by METTL3 plays a crucial role in promoting GC progression by reducing ferroptosis, through the modulation of the miR-27a-5p/BTF3 axis.
METTL3驱动的lncRNA FAM230B的m6A修饰通过调节miR-27a-5p/BTF3轴抑制胃癌的铁变态反应
我们之前的研究揭示了 lncRNA FAM230B 在胃癌(GC)中的凋亡抑制作用。但其在胃癌铁凋亡中的作用仍有待探索。本研究通过m6A定量、稳定性检测、MeRIP等方法检测了METTL3对FAM230B的m6A水平和RNA稳定性调控,并通过RIP和RNA牵引检测证实了它们之间的相互作用。流式细胞术、MDA 和 GSH 水平评估以及电子显微镜检测了铁变态反应的水平。基因表达通过实时定量 PCR、Western 印迹和免疫荧光进行检测。利用 TargetScan 预测了 miR-27a-5p 与 BTF3 的相互作用,并通过双荧光素酶检测进行了证实。在此观察到 GC 组织和细胞系中 METTL3 和 FAM230B 水平升高。证实METTL3与FAM230B RNA结合。此外,沉默 METTL3 会降低 FAM230B m6A 的水平和稳定性,从而导致 FAM230B 表达减少,miR-27a-5p 表达增加。敲除 FAM230B 有利于铁突变并增加 BTF3 的表达,而过表达 FAM230B 可减轻麦角新碱诱导的 GC 细胞铁突变。此外,研究还发现 BTF3 的过表达会抵消 miR-27a-5p 在 GC 细胞中促进铁变态反应的作用。总之,我们的研究表明,METTL3对FAM230B的m6A修饰通过调节miR-27a-5p/BTF3轴,在减少铁凋亡从而促进GC进展方面起着至关重要的作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Biochimica et biophysica acta. General subjects
Biochimica et biophysica acta. General subjects 生物-生化与分子生物学
CiteScore
6.40
自引率
0.00%
发文量
139
审稿时长
30 days
期刊介绍: BBA General Subjects accepts for submission either original, hypothesis-driven studies or reviews covering subjects in biochemistry and biophysics that are considered to have general interest for a wide audience. Manuscripts with interdisciplinary approaches are especially encouraged.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信