Ferroptosis, a newly discovered form of iron-dependent regulated cell death, has been implicated in traumatic brain injury (TBI). MiR-212-5p has previously been reported to be downregulated in extracellular vesicles following TBI. To investigate whether miR-212-5p is involved in the ferroptotic neuronal death in TBI mice, we first examined the accumulation of malondialdehyde (MDA) and ferrous ion, and the expression of ferroptosis-related molecules at 6 h, 12 h, 24 h, 48 h and 72 h following controlled cortical impact (CCI) in mice. There was a significant upregulation in the expression ofGpx4andAcsl4at 6 h,Slc7a11from 12 h to 72 h, andNox2andSat1from 6 h to 72 h post injury. Similarly, an upregulation in the expression of Gpx4 at 6 h, Nox2 from 6 h to 72 h, xCT from 12 h to 72 h, and Sat1 at 72 h after CCI was observed at the protein level. Interestingly, MDA and ferrous ion were increased whereas miR-212-5p was decreased in the CCI group compared to the sham group. Furthermore, we found that overexpression of miR-212-5p attenuated ferroptosis while downregulation of miR-212-5p promoted ferroptotic cell death partially by targeting prostaglandin-endoperoxide synthase-2 (Ptgs2) in HT-22 and Neuro-2a cell lines. In addition, administration of miR-212-5p in CCI mice significantly improved learning and spatial memory. Collectively, these findings indicate that miR-212-5p may protect against ferroptotic neuronal death in CCI mice partially by targeting Ptgs2.

Introduction: Despite advances in colorectal cancer (CRC) treatment, the chromosomal instability (CIN)-positive subgroup remains refractory to conventional therapies and immunotherapy. C2orf76, a poorly characterized gene identified as a CIN-linked candidate in CRC through bioinformatic analyses, has been associated with favorable prognosis and potential immune-related functions. However, its biological roles and underlying mechanisms in CRC remain largely unexplored. Methods: To investigate the role of C2orf76, we generated knockout (KO), overexpression (OE), and rescue (RE) cell lines in CRC models. We performed RNA-seq, RT-qPCR, and Western blotting to examine transcriptomic and protein-level changes. Functional assays included proliferation, migration, cisplatin sensitivity, pyroptosis (GSDME cleavage), cytokinesis-block micronucleus assay (CBMN) for CIN, and γ-H2AX foci detection for DNA double-strand breaks. Results: Unexpectedly, C2orf76 KO suppressed proliferation and migration, contradicting its favorable prognostic association and suggesting a complex regulatory role. RNA-seq revealed altered genes involved in chromosome segregation, cell cycle, platinum resistance, P53 signaling, and oncogenic pathways. RT-qPCR validated changes in chromosomal stability-related genes CDT1 and FOXM1, which were further verified by Western blotting. Further experiments showed that KO inhibited cisplatin-induced pyroptosis and cleavage of GSDME, whereas OE promoted migration and enhanced cisplatin sensitivity. The CBMN assay confirmed that C2orf76 significantly impacts chromosomal instability levels in CRC cells. C2orf76 KO cells also exhibited elevated γ-H2AX foci after cisplatin treatment. Rescue of C2orf76 expression reversed the KO-induced suppression of proliferation and migration, restored cisplatin sensitivity and pyroptosis, and reduced chromosomal instability. Discussion: These findings unveil a context-dependent duality of C2orf76: knockout increases CIN, reduces proliferation and migration, but promotes chemoresistance; overexpression enhances cisplatin-induced pyroptosis and drug sensitivity yet increases migration. Thus, C2orf76 establishes a functional link between CIN, cell death, and therapeutic response, positioning it as a tunable target that may require context-dependent modulation for effective CRC intervention. This study not only characterizes a previously understudied gene but also provides a mechanistic framework for understanding how CIN-associated genes can exert opposing effects on tumor progression and treatment outcome.