RBM15 facilitates laryngeal squamous cell carcinoma progression by regulating TMBIM6 stability through IGF2BP3 dependent.
RBM15 facilitates laryngeal squamous cell carcinoma progression by regulating TMBIM6 stability through IGF2BP3 dependent.
复制标题
DOI：
10.1186/s13046-021-01871-4
复制
发表时间：
2021-02-26
期刊：
影响因子：
--
通讯作者：
Sun Y
中科院分区：
文献类型：
--
作者：
Wang X;Tian L;Li Y;Wang J;Yan B;Yang L;Li Q;Zhao R;Liu M;Wang P;Sun Y
喉癌是头颈部肿瘤中死亡率最高的。RNA N6-甲基腺苷(M6A)是哺乳动物中含量最丰富、变化最大的一种核糖核酸。然而，喉鳞状细胞癌发生或发展的m6A调控机制仍然知之甚少。用m6A RNA甲基化定量试剂盒检测组织甲基化水平。M6A微阵列分析、mRNA转录序列分析和蛋白质组学检测RBM15、TMBIM6和IGF2BP3。采用免疫组织化学(IHC)、实时定量聚合酶链式反应(QRT-PCR)和免疫印迹法检测RBM15、TMBIM6和IGF2BP3在组织标本和细胞系中的表达。体外和体内检测了RBM15的生物学效应。通过RNA免疫沉淀(RIP)、甲基化RNA免疫沉淀测序(MERIP-seq)、RNase MAZF和荧光素酶报告实验验证了RBM15/IGF2BP3与TMBIM6的结合关系。用核糖核酸酶MAZF检测TMBIM6基因的甲基化位点。Hoechst染色证实细胞凋亡的变化。放线菌素D验证了TMBIM6的稳定性。喉鳞状细胞癌患者外周血中m6A基因甲基化水平显著升高。RBM15作为甲基转移酶的“写入者”，在喉癌组织中的表达显著升高，且与预后不良有关。在体内外，RBM15基因的敲除均可抑制喉鳞状细胞癌的增殖、侵袭、迁移和凋亡。在过量表达RBM15后，结果正好相反。从机制上讲，TMBIM6是RBM15介导的m6A修饰的下游靶点。此外，RBM15介导的M6A修饰TMBIM6mRNA通过依赖IGF2BP3来增强TMBIM6的稳定性。我们的结果揭示了RBM15和IGF2BP3在喉癌m6A甲基化修饰中的重要作用，从而确定了一种新的RNA调控机制。网上版载有补充材料，可在10.1186/s13046-021-01871-4查阅。
Laryngeal cancer has the highest mortality rate among head and neck tumours. RNA N6-methyladenosine (m6A) is the most plentiful and variable in mammalian mRNA. Yet, the m6A regulatory mechanism underlying the carcinogenesis or progression of LSCC remains poorly understood. The m6A RNA methylation quantification kit was used to detect tissue methylation levels. m6A microarray analysis, mRNA transcriptomic sequencing (mRNA-seq), and proteomics were used to determine RBM15, TMBIM6, and IGF2BP3. Immunohistochemical (IHC), quantitative real-time PCR (qRT-PCR) and Western blot were used to investigate RBM15, TMBIM6, and IGF2BP3 expression in tissue samples and cell lines. The biological effects of RBM15 were detected both in vitro and in vivo. The combination relationship between RBM15/IGF2BP3 and TMBIM6 was verified by RNA immunoprecipitation (RIP) assay, Methylated RNA immunoprecipitation sequencing (MeRIP-seq), RNase Mazf, and luciferase report assay. RNase Mazf was used to determine the methylation site on TMBIM6 mRNA. Hoechst staining assay was used to confirm the apoptotic changes. The actinomycin D verified TMBIM6 stability. The global mRNA m6A methylation level significantly increased in LSCC patients. RBM15, as a “writer” of methyltransferase, was significantly increased in LSCC and was associated with unfavorable prognosis. The knockdown of RBM15 reduced the proliferation, invasion, migration, and apoptosis of LSCC both in vitro and in vivo. The results were reversed after overexpressing RBM15. Mechanically, TMBIM6 acted as a downstream target of RBM15-mediated m6A modification. Furthermore, RBM15-mediated m6A modification of TMBIM6 mRNA enhanced TMBIM6 stability through IGF2BP3-dependent. Our results revealed the essential roles of RBM15 and IGF2BP3 in m6A methylation modification in LSCC, thus identifying a novel RNA regulatory mechanism. The online version contains supplementary material available at 10.1186/s13046-021-01871-4.
登录
查看更多内容
影响因子：
--
作者：
Sun C;Han C;Wang P;Jin Y;Sun Y;Qu L
通讯作者：
Qu L
影响因子：
4.7
作者：
Bai, Yang;Yang, Chunxing;Zhang, Yi
通讯作者：
Zhang, Yi
影响因子：
254.7
作者：
Siegel, Rebecca;Ma, Jiemin;Jemal, Ahmedin
通讯作者：
Jemal, Ahmedin
影响因子：
50.3
作者：
Li Z;Weng H;Su R;Weng X;Zuo Z;Li C;Huang H;Nachtergaele S;Dong L;Hu C;Qin X;Tang L;Wang Y;Hong GM;Huang H;Wang X;Chen P;Gurbuxani S;Arnovitz S;Li Y;Li S;Strong J;Neilly MB;Larson RA;Jiang X;Zhang P;Jin J;He C;Chen J
通讯作者：
Chen J
胰岛素样生长因子 2 mRNA 结合蛋白 2 通过稳定 GLUT1 mRNA 促进胰腺导管腺癌中的有氧糖酵解和细胞增殖
影响因子：
3.7
作者：
Huang, Shan;Wu, Zheng;Hao, Linlin
通讯作者：
Hao, Linlin