Nε-Carboxymethyl-Lysine Drives Centrosome Amplification and Hepatocarcinogenesis via Pathological Dysregulation of the PLK1-CEP20-CCDC116 Axis.

Thermal processing of foods generates hepatotoxic dietary advanced glycation end products (dAGEs), with Nε-carboxymethyl-lysine (CML) serving as a primary carcinogen. Chronic exposure to physiologically relevant CML concentrations (25 μg/mL) induced centrosome amplification (CA)─a hallmark of genomic instability─in human hepatocytes (THLE2/THLE3), driving malignant transformation marked by 5-fold increased proliferation, 3-fold enhanced migration, and murine tumorigenicity. Critically, CML-treated cells exhibited 8-10-fold higher CA frequencies, with CA-positive subpopulations showing elevated oncogenicity. Mechanistically, CML activated the PLK1-CEP20-CCDC116 axis through EGR1-mediated PLK1 transcriptional upregulation and DNMT3B-suppression-driven CEP20 promoter hypomethylation, culminating in the PLK1-dependent phosphorylation of CEP20 at Ser46. This post-translational modification enabled high-affinity CCDC116 binding and centrosomal recruitment, driving centriole overduplication. Critically, both the CEP20-S46A mutation and CCDC116 knockdown abolished centrosomal localization and CA induction. These findings establish exogenous CML as a direct inducer of organelle-level carcinogenesis and provide molecular targets for developing food safety interventions against thermally processed food toxicity.