N6-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 N6-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, m6A sequencing and m6A methylated RNA immuno-precipitation quantitative reverse-transcription polymerase chain reaction, we revealed the profile of METTL3-mediated m6A modification in BCa cells for the first time. AF4/FMR2 family member 4 (AFF4), two key regulators of NF-κB pathway (IKBKB and RELA) and MYC were further identified as direct targets of METTL3-mediated m6A modification. In addition, we showed that besides NF-κB, 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-κB/MYC signaling network operated by METTL3-mediated m6A modification and provided insight into the mechanisms of BCa progression.
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The authors declare that all relevant data are available within the article and its Supplementary information files or from the corresponding author upon reasonable request. The m6A-sequencing and RNA-sequencing datasets have been submitted to the NCBI database under the accession number PRJNA498900, SAMN10337857 and SAMN10337858.
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This work was supported by the National Natural Science Foundation of China (81872313 and 81672776 to YL, 81802391 to QG, 31501838 to XH Z), and Anhui Provincial Natural Science Foundation (1808085QH266 to QG)
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Cheng, M., Sheng, L., Gao, Q. et al. The m6A methyltransferase METTL3 promotes bladder cancer progression via AFF4/NF-κB/MYC signaling network. Oncogene 38, 3667–3680 (2019). https://doi.org/10.1038/s41388-019-0683-z
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