NAT10 promotes colon cancer progression by enhancing predicted N4-acetylcytidine modification of Notch2 mRNA.

BACKGROUND

Colon cancer is a prevalent malignant tumor with high mortality, and RNA modifications play critical roles in its progression. N-acetyltransferase 10 (NAT10)-mediated N4-acetylcytidine (ac4C) modification has emerged as a key epigenetic regulator in gastrointestinal cancers, but its role and mechanism in colon cancer remain unclear. This study investigated whether NAT10 regulates Notch2 via predicted ac4C modification to affect colon cancer progression.

MATERIALS AND

METHODS

NAT10 and Notch2 expression was detected in 15 pairs of colon cancer and adjacent normal tissues. Colon cancer cell lines (HCT116, SW480) and normal colonic epithelial NCM460 cells were used. NAT10 overexpression/knockdown, NAT10 inhibitor Remodelin treatment, rescue experiments in both cell lines, and in vivo xenograft models were performed. CCK-8, Transwell, flow cytometry, TUNEL, acRIP-qPCR, RIP-qPCR, dual-luciferase reporter (WT/MUT), and mRNA stability assays were applied.

RESULTS

NAT10 and Notch2 were significantly upregulated in colon cancer tissues and cells, with a positive correlation trend (r = 0.3571, P = 0.1913). NAT10 knockdown inhibited proliferation, migration, and promoted apoptosis; overexpression exerted opposite effects. Remodelin inhibited NAT10 activity and downregulated Notch2 in a dose-dependent manner. Mechanistically, NAT10 bound to Notch2 mRNA, enhanced its predicted ac4C modification and stability. The predicted 3'UTR ac4C site was critical for NAT10-mediated regulation. Notch2 overexpression reversed NAT10 knockdown effects in both HCT116 and SW480 cells.In vivo subcutaneous xenograft models further confirmed that NAT10 knockdown significantly inhibits colon cancer tumor growth in mice, while Notch2 overexpression effectively rescues this inhibitory effect, supporting the oncogenic role of the NAT10/ac4C/Notch2 axis in vivo.

CONCLUSION

NAT10 promotes colon cancer progression in an enzymatic activity‑dependent manner by enhancing predicted ac4C modification of Notch2 mRNA to stabilize its expression. This study reveals a novel NAT10/ac4C/Notch2 regulatory axis and provides potential therapeutic targets for colon cancer.