FUS and METTL3 collaborate to regulate RNA maturation, preventing unfolded protein response and promoting gastric cancer progression.

Publisher: Springer-Verlag Italia Country of Publication: Italy NLM ID: 100973405 Publication Model: Electronic Cited Medium: Internet ISSN: 1591-9528 (Electronic) Linking ISSN: 15918890 NLM ISO Abbreviation: Clin Exp Med Subsets: MEDLINE

Original Publication: Milano, Italy: Springer-Verlag Italia, c2001-

Stomach Neoplasms*/genetics ; Stomach Neoplasms*/pathology ; Stomach Neoplasms*/metabolism ; RNA-Binding Protein FUS*/genetics ; RNA-Binding Protein FUS*/metabolism ; Methyltransferases*/metabolism ; Methyltransferases*/genetics ; Mice, Nude* ; Unfolded Protein Response*; Animals ; Humans ; Mice ; Apoptosis ; Cell Line, Tumor ; Endoplasmic Reticulum Stress ; Gene Expression Regulation, Neoplastic ; Disease Progression ; Alternative Splicing ; Disease Models, Animal ; Female ; Male

FUS-mediated alternative splicing and METTL3-regulated RNA methylation play crucial roles in RNA processing. The purpose of this study was to investigate the interactive roles of FUS and METTL3 in gastric cancer (GC) progression. RNA sequencing data were obtained from the TCGA-STAD dataset. Differentially expressed genes (DEGs) were analyzed across groups stratified by the medians of FUS, METTL3, and NEAT1, respectively. Endoplasmic reticulum (ER) stress markers PERK, IRE1, pIRE1, Bip, and CHOP, as well as related apoptosis stress markers PARP, cleaved-PARP, (Cleaved) Caspase 7, and (Cleaved) Caspase 3, were assessed through western blotting. Alternative splicing and N6-methyladenosine (m(6)A) methylation of specific genes were detected with MeRIP-PCR. Finally, in vivo experiments were conducted using nude mice bearing sh-FUS-transfected HGC27 xenograft tumors. FUS and METTL3 expression levels were elevated in GC tissues. A significant overlap of DEGs was observed between the FUS- and METTL3-stratified groups. These overlapping DEGs were predominantly enriched in mRNA processing and protein processing in the ER. ER stress and apoptosis were induced by sh-FUS or sh-METTL3, which was further enhanced by ER stress inducer tunicamycin in both MKN45 and HGC27 cells. Similarly, DEGs for NEAT1 high- and low-expressed groups were enriched in protein processing in the ER and spliceosome. To a lesser extent, ER stress was also induced by sh-NEAT1 and enhanced by tunicamycin in HGC27 cells. Furthermore, sh-FUS or sh-METTL3 influenced alternative splicing and methylation of specific mRNAs, including FUS, NEAT1, PCNA, MCM2, and BIRC5. Tumor progression was inhibited by sh-FUS in mice, and ER stress and apoptosis were induced, which were further enhanced by tunicamycin. FUS and METTL3 collaborate to facilitate RNA maturation. Inhibiting FUS or METTL3 promoted ER stress and apoptosis and inhibited progression in GC.
Competing Interests: Declarations. Conflict of interest: The authors declare no competing interests. Ethical approval: Animal experiments were conducted under the approval of the Ethics Committee of General Hospital of Ningxia Medical University (approval no.: KYLL-2021–766).
(© 2024. The Author(s).)

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2022BEG03154 the Key Research and Development Project of Ningxia Science and Technology Department

Keywords: Alternative splicing; FUS; METTL3; NEAT1; RNA processing; Unfolded protein response

0 (RNA-Binding Protein FUS)
EC 2.1.1.- (Methyltransferases)
EC 2.1.1.62 (METTL3 protein, human)
0 (FUS protein, human)

Date Created: 20241221 Date Completed: 20241221 Latest Revision: 20241221

20241222

10.1007/s10238-024-01525-7

39708203