N6-methyladenosine modification of ITGA6 mRNA promotes the development and progression of bladder cancer
N6-methyladenosine modification of ITGA6 mRNA promotes the development and progression of bladder cancer
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ITGA6 mRNA N-6-甲基腺苷修饰促进膀胱癌的发生和进展
DOI：
10.1016/j.ebiom.2019.07.068
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发表时间：
2019-09-01
期刊：
影响因子：
11.1
通讯作者：
Ji, Weidong
中科院分区：
文献类型：
--
作者：
Jin, Huan;Ying, Xiaoling;Ji, Weidong
背景：越来越多的证据表明，mRNA的N - 6 -甲基腺苷（m(6)A）修饰在各种癌症中起着关键作用。然而，m(6)A在膀胱癌（BC）中的生物学功能和调控机制尚未完全明确。
方法：我们进行了细胞表型分析，并建立了体内小鼠异种移植模型，以评估m(6)A修饰的ITGA6对膀胱癌生长和进展的影响。采用甲基化RNA免疫沉淀（MeRIP）、RNA免疫沉淀以及荧光素酶报告基因和诱变分析来确定m(6)A修饰的ITGA6的作用机制。进行免疫组化分析以评估膀胱癌患者中METTL3和ITGA6表达之间的相关性。
研究结果：我们发现m(6)A“书写器”METTL3和“橡皮擦”ALKBH5通过调节膀胱癌细胞中ITGA6的表达来改变细胞黏附。此外，在人膀胱癌组织中，ITGA6的上调与METTL3表达的增加相关，且患者中ITGA6较高的表达预示着较低的生存率。从机制上讲，m(6)A在ITGA6转录本中高度富集，ITGA6 mRNA 3'非翻译区（3'UTR）的m(6)A甲基化增加通过m(6)A“阅读器”YTHDF1和YTHDF3的结合促进ITGA6 mRNA的翻译。抑制ITGA6可导致膀胱癌细胞在体外和体内的生长和进展减缓。此外，在METTL3缺失的细胞中过表达ITGA6可部分恢复膀胱癌的黏附、迁移和侵袭表型。
解读：我们的研究结果证明了m(6)A修饰的ITGA6的致癌作用，并展示了其在膀胱癌发生和进展中的调控机制，从而确定了一个膀胱癌的潜在治疗靶点。
基金：本研究由中国国家自然科学基金（81772699，81472999）资助。（c）2019作者。由爱思唯尔出版公司出版。这是一篇在知识共享署名 - 非商业性使用 - 禁止演绎（CC BY - NC - ND）许可协议下的开放获取文章（http://creativecommons.org/licenses/by - nc - nd/4.0/）
Background: Accumulating evidence has revealed the critical roles of N-6-methyladenosine (m(6)A) modification of mRNA in various cancers. However, the biological function and regulation of m(6)A in bladder cancer (BC) are not yet fully understood.Methods: We performed cell phenotype analysis and established in vivo mouse xenograft models to assess the effects of m(6)A-modified ITGA6 on BC growth and progression. Methylated RNA immunoprecipitation (MeRIP), RNA immunoprecipitation and luciferase reporter and mutagenesis assays were used to define the mechanism of m(6)A-modified ITGA6. Immunohistochemical analysis was performed to assess the correlation between METTL3 and ITGA6 expression in bladder cancer patients.Findings: We show that the m(6)A writer METTL3 and eraser ALKBH5 altered cell adhesion by regulating ITGA6 expression in bladder cancer cells. Moreover, upregulation of ITGA6 is correlated with the increase in METTL3 expression in human BC tissues, and higher expression of ITGA6 in patients indicates a lower survival rate. Mechanistically, m(6)A is highly enriched within the ITGA6 transcripts, and increased m(6)A methylations of the ITGA6 mRNA 3'UTR promotes the translation of ITGA6 mRNA via binding of the m(6)A readers YTHDF1 and YTHDF3. Inhibition of ITGA6 results in decreased growth and progression of bladder cancer cells in vitro and in vivo. Furthermore, overexpression of ITGA6 inMETTL3-depleted cells partially restores the BC adhesion, migration and invasion phenotypes.Interpretation: Our results demonstrate an oncogenic role of m(6)A-modified ITGA6 and show its regulatory mechanisms in BC development and progression, thus identifying a potential therapeutic target for BC.Fund: This work was supported by National Natural Science Foundation of China (81772699, 81472999). (c) 2019 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).