Background: Tumor cells tend to utilize glycolysis rather than aerobic respiration even under aerobic conditions. OVOL2, an inhibitory C2H2 zinc finger transcription factor, is a potential tumor suppressor in cancers. However, the association between OVOL2 and tumor energy metabolism is unknown. Methods: Western blotting was used to determine the expression of OVOL2 in different non-small cell lung cancer (NSCLC) cell lines and mouse models. The metabolic parameters in NSCLC cells following overexpression or knockdown OVOL2 were examined. To define the mechanism by which OVOL2 regulates aerobic glycolysis, interacting protein of OVOl2 and downstream molecular events were identified by luciferase assay and co-immunoprecipitation. We documented the regulatory mechanism in mouse xenograft models. Finally, clinical relevance of OVOL2, NF-κB signaling and GLUT1 was measured by immunostaining. Results: OVOL2 is downregulated in NSCLC and overexpression of OVOL2 inhibits the survival of cancer cells. Moreover, OVOL2 directly binds to P65 and inhibits the recruitment of P300 but facilitates the binding of HDAC1 to P65, which in turn negatively regulates NF-κB signaling to suppress GLUT1 translocation and glucose import. In contrast, OVOL2 expression is negatively regulated by NF-κB signaling in NSCLC cells via the ubiquitin–proteasome pathway. Re-expression of OVOL2 significantly compromise NF-κB signaling-induced GLUT1 translocation, aerobic glycolysis in NSCLC cells and mouse models. Immunostaining revealed inverse correlations between the OVOL2 and phosphorylated P65 levels and between the OVOL2 and membrane GLUT1 levels, and a strong correlation between the phosphorylated P65 and membrane GLUT1 levels. Conclusions: These results suggest a regulatory circuit linking NF-κB and OVOL2, which highlights the role of NF-κB signaling and OVOL2 in the modulation of glucose metabolism in NSCLC.

Acute myocardial infarction (AMI) benefits from biomarkers and assays that support early triage. Cardiac troponin T (cTnT) is widely used, but may be less informative early after ischemia and can be elevated in non-AMI conditions. Circulating miR-499 has been reported to rise early after myocardial injury, yet miRNA detection is complicated by short target length and PAM constraints in many CRISPR-Cas12a designs. We developed a PAM-independent, one-tube isothermal RT–RPA–Cas12a platform (RCNPM) for miR-499 and evaluated its analytical performance and clinical discrimination, with routine clinical cTnT analyzed in parallel. RCNPM integrates stem-loop reverse transcription, recombinase polymerase amplification (RPA), and Cas12a trans-cleavage fluorescence readout at 37 °C in a single vessel. Primers and reaction components were screened/optimized, and specificity was assessed using non-target miRNAs. Clinical evaluation was performed in a single-center prospective cohort (n = 150; 75 AMI meeting ESC criteria, 75 controls) with random coding and blinded miR-499 testing. RCNPM was benchmarked against RT–qPCR (Spearman correlation; Bland–Altman). Serum cTnT was obtained from routine hospital immunoassay testing and was not measured within the RCNPM reaction. ROC analyses were performed with 95% CIs. RCNPM detected miR-499 down to 1 aM and showed no detectable signal in the non-target miRNA panel. The RT–RPA–Cas12a step required ~ 30 min (workflow including extraction ~ 45–60 min). In the cohort, RCNPM agreed with RT–qPCR (Spearman r = 0.96, p < 0.001) with minimal mean bias. In this dataset, ROC analyses yielded AUC = 1.00 (95% CI 1.00–1.00) for miR-499 (RCNPM and RT–qPCR) and routine cTnT. RCNPM enables PAM-independent, one-tube isothermal miR-499 detection with strong agreement to RT–qPCR in a single-center cohort. External validation in larger, multi-center and more heterogeneous disease-control cohorts is warranted. The online version contains supplementary material available at 10.1186/s12967-026-08116-y.