The m6A methyltransferase METTL3 promotes bladder cancer progression via AFF4/NF-κB/MYC signaling network
The m6A methyltransferase METTL3 promotes bladder cancer progression via AFF4/NF-κB/MYC signaling network
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m(6)A 甲基转移酶 METTL3 通过 AFF4/NF-kappa B/MYC 信号网络促进膀胱癌进展
DOI：
10.1038/s41388-019-0683-z
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发表时间：
2019-05-09
期刊：
影响因子：
8
通讯作者：
Li, Yang
中科院分区：
文献类型：
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作者：
Cheng, Maosheng;Sheng, Lu;Li, Yang
N-6-甲基腺苷（m6 A）是真核生物信使RNA（mRNAs）中最丰富的修饰形式，在许多生物过程中发挥重要作用。然而，其在膀胱癌（BCa）中的功能仍然难以捉摸。在这里，我们发现，甲基转移酶样3（胃L3），一个主要的RNA N-6-腺苷甲基转移酶，在人类BCa显着上调。敲除胃L3显著降低了BCa细胞的增殖、侵袭和体外存活以及体内致瘤性。另一方面，胃L3的过表达显著促进BCa细胞的生长和侵袭。通过转录组测序、m（6）A测序和m（6）A甲基化RNA免疫沉淀定量逆转录聚合酶链反应，我们首次揭示了胃L3介导的m（6）A修饰在BCa细胞中的分布。NF-κ B通路的两个关键调节因子AF 4/FMR 2家族成员4（AFF 4）和MYC被进一步鉴定为胃L3介导的m（6）A修饰的直接靶点。此外，我们发现，除了NF-κ B，AFF 4结合到MYC的启动子，并促进其表达，这意味着一个新的多级调控网络下游的胃L3。我们的研究结果揭示了由胃L3介导的m6 A修饰操作的AFF 4/NF-κ B/MYC信号网络，并为BCa进展的机制提供了见解。
N-6-methyladenosine (m6A) is the most abundant modification in eukaryotic messenger RNAs (mRNAs), and plays important roles in many bioprocesses. However, its functions in bladder cancer (BCa) remain elusive. Here, we discovered that methyltransferase-like 3 (METTL3), a major RNA N-6-adenosine methyltransferase, was significantly up-regulated in human BCa. Knockdown of METTL3 drastically reduced BCa cell proliferation, invasion, and survival in vitro and tumorigenicity in vivo. On the other hand, overexpression of METTL3 significantly promoted BCa cell growth and invasion. Through transcriptome sequencing, m(6)A sequencing and m(6)A methylated RNA immuno-precipitation quantitative reverse-transcription polymerase chain reaction, we revealed the profile of METTL3-mediated m(6)A modification in BCa cells for the first time. AF4/FMR2 family member 4 (AFF4), two key regulators of NF-kappa B pathway (IKBKB and RELA) and MYC were further identified as direct targets of METTL3-mediated m(6)A modification. In addition, we showed that besides NF-kappa B, AFF4 binds to the promoter of MYC and promotes its expression, implying a novel multilevel regulatory network downstream of METTL3. Our results uncovered an AFF4/NF-kappa B/MYC signaling network operated by METTL3-mediated m6A modification and provided insight into the mechanisms of BCa progression.