# Identification and experimental verification of necroptosis-related prognostic gene signature and characterization of tumor immune infiltration in lung squamous cell carcinoma

## TL;DR

This study identifies a three-gene signature linked to necroptosis in lung squamous cell carcinoma, which helps predict patient outcomes and immune response patterns.

## Contribution

A novel three-gene necroptosis-related signature is identified and validated for prognosis and immune infiltration in LUSC.

## Key findings

- Three necroptosis-related genes (CAMK2A, CHMP4C, and PYGB) were found to be strongly associated with patient prognosis in LUSC.
- The three-gene signature was validated to have moderate predictive accuracy for survival outcomes in LUSC patients.
- The genes correlate with immune infiltration, immune checkpoint activity, tumor mutation burden, and microsatellite instability.

## Abstract

Lung squamous cell carcinoma (LUSC) is a common and aggressive malignancy. Necroptosis, a regulated mode of cell death, has been implicated in tumor immunity and oncogenic processes, yet the mechanistic involvement of necroptosis-related genes (NRGs) in LUSC pathogenesis remains unclear, necessitating systematic evaluation of their biological and clinical relevance.

Clinical and transcriptomic data of LUSC patients from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) were subjected to integrative analyses. Screening of the Kyoto Encyclopedia of Genes and Genomes (KEGG) database identified 159 NRGs, among which 35 differentially expressed NRGs (DENRGs) were associated with necroptosis, apoptosis, and immune signaling pathways. Cox regression combined with Least Absolute Shrinkage and Selection Operator (LASSO) analysis yielded three NRGs (CAMK2A, CHMP4C, and PYGB) strongly associated with patient prognosis. Based on these genes, a prognostic model was constructed to stratify patients into high- and low-risk subgroups with distinct survival patterns. External dataset validation demonstrated moderate predictive accuracy. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and immunohistochemistry (IHC) confirmed abnormal expression of the three genes in LUSC tissues. Additional analyses revealed correlations of these NRGs with immune infiltration, immune checkpoint activity, tumor mutation burden (TMB), and microsatellite instability (MSI).

A three-gene NRG signature was identified as a prognostic marker in LUSC. These genes appear to influence disease progression and the immune microenvironment, highlighting their potential as therapeutic targets and as a foundation for further investigation.