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N6-methyladenosine demethylase ALKBH5 suppresses malignancy of esophageal cancer by regulating microRNA biogenesis and RAI1 expression

Oncogene volume 40, pages 5600–5612 (2021)Cite this article

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Abstract

N6-Methyladenosine (m6A) is the most prevalent epigenetic RNA modification and is vital in regulating malignancies. The roles of m6A modifiers on noncoding RNAs have not been fully investigated in esophageal cancer. By screening all m6A modifiers, ALKBH5 was the most potent member related to patient outcomes and suppressing esophageal cancer malignancy in cell and animal models. It demethylated pri-miR-194-2 and inhibited miR-194-2 biogenesis through an m6A/DGCR8-dependent manner. RAI1, previously considered as a circadian clock transcriptional regulator, was the main target of miR-194-2. It enhanced transcription of Hippo pathway upstream genes by binding to their 3′UTR and suppressed YAP/TAZ nuclear translocation. The ALKBH5/miR-194-2/RAI1 axis was also validated in clinical samples. In addition, the increased malignancy by low ALKBH5 was abolished by the YAP inhibitor verteporfin. Our findings uncover a critical role of ALKBH5 in miRNAs biogenesis and provide novel insight for developing treatment strategies in esophageal cancer.

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Fig. 1: ALKBH5 correlates with a good prognosis in esophageal cancer patients.
Fig. 2: ALKBH5 impedes cell growth and motility of esophageal cancer in vitro and in vivo.
Fig. 3: ALKBH5 suppresses the biogenesis of miR-194-2 and miR-532 clusters.
Fig. 4: ALKBH5-mediated tumor suppression and miRNA biogenesis rely on m6A modification.
Fig. 5: miR-194-2 is onco-promoting in esophageal cancer and counteracts ALKBH5 function.
Fig. 6: RAI1 is a target gene of ALKBH5/miR-194-2.
Fig. 7: RAI1 correlates with outcomes of esophageal cancer patients and hinders tumor progression in vitro.
Fig. 8: ALKBH5/RAI1 regulates the Hippo pathway and sensitivity to verteporfin.

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Acknowledgements

Alexandra Marshall from Marshall Medical Communications provided editorial assistance during manuscript preparation. This study was supported by the National Natural Science Foundation of China (81773228 and 81972850 to Y.C.; 81601999 to L.Z.); Postdoctoral Science Foundation of China (2018M640637 to L.Z.), and Special Fund for Taishan Scholar Project (ts20190973 to Y.C.).

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Author notes

  1. These authors contributed equally: Pengxiang Chen and Song Li.

Authors and Affiliations

  1. Department of Radiation Oncology, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, People’s Republic of China

    Pengxiang Chen, Wei Zhou, Yuchen Liu, Lin Zhang & Yufeng Cheng

  2. Laboratory of Basic Medical Sciences, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, People’s Republic of China

    Pengxiang Chen

  3. Department of Medical Oncology, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, People’s Republic of China

    Song Li

  4. Department of General Medicine, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, People’s Republic of China

    Ke Zhang

  5. Thoracic Surgery Department, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, People’s Republic of China

    Renchang Zhao

  6. Department of Urology, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, People’s Republic of China

    Jianfeng Cui

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Contributions

Y.C. and L.Z. conceived and designed the study. P.C. performed most of the in vitro and in vivo experiments. S.L. performed the in silico studies and helped with the in vivo studies. R.Z., J.C., W.Z., and Y.L. helped with the clinical studies. Y.C., L.Z., P.C., and S.L. analyzed data and drafted the manuscript. P.C., S.L., K.Z., R.Z., and Y.L. supplemented the experiments according to the reviewers’ comments. All authors read and approved the final manuscript.

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Correspondence to Lin Zhang or Yufeng Cheng.

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Chen, P., Li, S., Zhang, K. et al. N6-methyladenosine demethylase ALKBH5 suppresses malignancy of esophageal cancer by regulating microRNA biogenesis and RAI1 expression. Oncogene 40, 5600–5612 (2021). https://doi.org/10.1038/s41388-021-01966-4

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