Demethylase ALKBH5 suppresses invasion of gastric cancer via PKMYT1 m6A modification

Molecular Cancer

Jiao Liu¹  Chunli Gong²  Yufeng Xiao²  Yu Huang²  Yiyang Hu²  Yang Chen²  Qiang Luo²  Zhibin Li²  Sumin Wang²  Shiming Yang²  Yu Hou³ 

[1]Department of Endoscope, General Hospital of Northern Theater Command, 110016, Shenyang, Liaoning, China

[2]Department of Gastroenterology, Xinqiao Hospital, Third Military Medical University, 400037, Chongqing, China

[3]Department of Hematology, Southwest Hospital, Third Military Medical University, 400038, Chongqing, China

[4]Institute of Life Sciences, Chongqing Medical University, 400016, Chongqing, China

关键词: ALKBH5;    Invasion;    Metastasis;    Demethylase activity;    PKMYT1;    Gastric cancer;   

DOI  :  10.1186/s12943-022-01522-y

来源: Springer

【 摘 要 】

BackgroundGastric cancer (GC) is one of the most pernicious tumors that seriously harm human healthcare. GC metastasis is one of the prime cause of failed cancer treatment, but correlation between N6-methyladenosine (m6A) and GC metastasis was less reported.MethodsMethylated RNA immunoprecipitation sequencing (MeRIP-seq) of GC tissues was conducted. Quantitative real-time PCR (qRT-PCR), western blotting and immunohistochemistry (IHC) were taken to determine the expression of ALKBH5 in GC tissues and cell lines. RNA-seq together with MeRIP-qRT-PCR was used to screen the target gene of ALKBH5. RNA pulldown, mass spectrometry and RNA immunoprecipitation (RIP) were used to search the “reader” protein of target gene. The mechanism was also validated via a tail vein injection method for lung metastasis model.ResultsDecreased expression of ALKBH5 was detected in GC samples, and it was correlated with clinical tumor distal metastasis and lymph node metastasis. ALKBH5 interference promoted metastasis of GC cells and this effect was closely related to the demethylase activity of ALKBH5. PKMYT1, as a downstream target of ALKBH5, promoted invasion and migration in GC. Caused by ALKBH5 knockdown or its demethylase activity mutation, upregulated expression of PKMYT1 indicated that ALKBH5 modulates expression of PKMYT1 in an m6A-dependent manner. IGF2BP3 helped stabilize the mRNA stability of PKMYT1 via its m6A modification site.ConclusionsThis study established an ALKBH5-PKMYT1-IGF2BP3 regulation system in metastasis, representing a new therapeutic target for GC metastasis.

【 授权许可】

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