NAT10 promotes gastric cancer metastasis via N4-acetylated COL5A1

Gastric cancer (GC) is among the most prevalent gastrointestinal malignancies. The occurrence of local deep infiltration or distant metastasis in GC is commonly associated with weak treatment and poor prognosis. 1 Although, N4-Acetylcytidine (ac4C) represents one of the extensive chemical modifications in mRNAs that plays a pivotal role in modulating mRNA stability and the mRNA translation process (Fig. 1b ). However, the role of mRNA ac4C modification in disease remains unclear. 2 As the only known ac4C “writer” protein, NAT10 is thought to have critical effects in tumor metastasis and tumor cell epithelial-to-mesenchymal transition (EMT). Here, we report a novel mechanism of NAT10-mediated mRNA ac4C modification regulating gastric cancer metastasis and EMT. Fig. 1 NAT10 promotes gastric cancer progression through mRNA ac4C modification. a Graphical summary of this article. b The chemical structure of ac4C. c The expression of NAT10 in the TCGA STAD dataset. d High NAT10 expression was significantly associated with a shorter OS. e The ac4C significant consensus sequence motif was identified based on the acRIP-seq and RIP-seq analyses, and the significant consensus sequence of NAT10 bound and interacted. f KEGG pathway enrichment analysis and acRIP-seq analysis identified the enriched pathway of ac4C modification down-regulated genes after NAT10 silencing. g Overlapping the potential target bound and interacted with NAT10 in focal adhesion and ECM-receptor interaction pathway by the acRIP-seq and RIP-seq analysis. h Attenuation of the NAT10 diminishes the ac4C modification genome of COL5A1 mRNA visual result compared using the acRIP-seq (colored in deep blue). The visual genome result of COL5A1 mRNA bound and interacted with NAT10 (colored in wathet blue). i The regulatory role of NAT10 on COL5A1 ac4C in SGC-7901 and MGC-803 cells confirmed by the acRIP-qPCR assay. j The bound and interacted relationship between NAT10 and COL5A1 mRNA in SGC-7901 and MGC-803 cells confirmed using the RIP-qPCR assay. k , l Western blotting and qRT-PCR analyses of COL5A1 in shNAT10 or NAT10 overexpressing MGC-803 and SGC-7901 cells. m Wild-type (WT) or mutant COL5A1 cells were transfected with pmirGLO-COL5A1 reporter, respectively. n The transcriptional level of wild-type COL5A1, but not the mutation, significantly decreased in the NAT10-knockdown cells, and significantly increased in the NAT10 overexpressing cells. o The impression of NAT10 on COL5A1 mRNA stability confirmed by the RNA decay assay. p The impression of NAT10 on COL5A1 mRNA translation efficiency confirmed Full size image