Expression of the NLRP12/NF-κB/iNOS axis in lung tissues across stages of non-small cell lung cancer and its mechanism in mediating immune cell regulation
TL;DR
The NLRP12/NF-κB/iNOS/Treg axis remodels the TIME to foster tumor progression, offering a novel target and experimental rationale for precision diagnosis and therapy in LUSC.
Abstract
Background Patients with lung squamous cell carcinoma (LUSC), a subtype of non-small cell lung cancer (NSCLC), face a dismal prognosis. Immune evasion driven by an imbalanced tumor immune microenvironment (TIME) and an abnormal enrichment of regulatory T cells (Tregs) is a central pathogenic mechanism, yet the regulatory pathways involved remain elusive. This study aims to investigate the expression profile and immunomodulatory mechanism of the NLR family pyrin domain containing 12 (NLRP12)/nuclear factor kappa B subunit 1 (NF-κB)/inducible nitric oxide synthase (iNOS) axis in LUSC, and to identify potential prognostic biomarkers and therapeutic targets. Methods We integrated bioinformatics analysis from databases such as gene expression profiling interactive analysis (GEPIA) and Kaplan-Meier Plotter with experimental validation in 48 clinical LUSC samples. Complementary in vitro models, including CD4+ T cell overexpression/silencing, pathway intervention, and co-culture assays, were employed. Mechanisms were verified using western blotting (WB), quantitative real-time polymerase chain reaction (qPCR), and flow cytometry. Results NLRP12 was highly expressed in LUSC and correlated with shortened overall survival (OS). Its expression showed a negative correlation with p-NF-κB and iNOS levels during tumor progression. Functionally, NLRP12 promoted the differentiation of CD4+ T cells into Tregs by inhibiting NF-κB phosphorylation and subsequent iNOS expression. The interleukin (IL)-10/transforming growth factor beta 1 (TGF-β) secreted by these NLRP12-driven Tregs enhanced the proliferation and invasion of LUSC cells in vitro, an effect reversed by neutralizing antibodies. Conclusions NLRP12 serves as a specific prognostic biomarker in LUSC. The NLRP12/NF-κB/iNOS/Treg axis remodels the TIME to foster tumor progression, offering a novel target and experimental rationale for precision diagnosis and therapy in LUSC.