MACF1 Promotes Colorectal Cancer Progression via ATF4-Mediated ASNS Regulation

Methods

MACF1 expression was assessed in CRC tissues and cell lines (HCT116, HT29, SW620, Caco2, LoVo) via immunohistochemistry and qPCR. Functional assays—including proliferation, colony formation, wound healing, Transwell migration, and cell-cycle analysis—were performed following MACF1 knockdown and overexpression. In vivo tumor growth was evaluated using a subcutaneous xenograft model.

RNA-seq screening identified ASNS as a downstream effector, and the MACF1–ATF4–ASNS relationship was explored through ChIP, immunofluorescence, nuclear fractionation, and Co-IP. Results: MACF1 was significantly elevated in CRC tissues and cells versus controls (p < 0.05). MACF1 knockdown suppressed proliferation, colony formation, migration, and xenograft tumor growth, while overexpression promoted aggressive phenotypes (p < 0.05).

Mechanistically, ASNS was identified as a critical MACF1 target, with ATF4 serving as a transcriptional mediator linking MACF1 to ASNS regulation. Conclusion: Our findings demonstrate that MACF1 promotes CRC cell growth and migration, at least in part, via ATF4-associated ASNS regulation and asparagine metabolism. This work establishes a novel functional bridge between cytoskeletal remodeling and amino acid metabolic reprogramming in CRC, offering potential insights for future metabolism-oriented therapeutic strategies.