CELL INTELLIGENCE · 肿瘤细胞治疗研究

Recent research has identified pyroptosis as a distinct form of programmed cell death that is strongly correlated with tumor outcomes. However, its value as a prognostic biomarker in clear cell renal cell carcinoma (ccRCC) is not yet fully established.

Differential expression of pyroptosis-related genes between ccRCC and normal tissues was identified using the DESeq2 R package. Next, univariate and least absolute shrinkage and selection operator (LASSO) Cox regression analyses using data from The Cancer Genome Atlas cohort were applied to develop a prognostic model. Univariate and multivariate prognostic analyses were conducted to determine whether the risk score could independently predicted patient prognosis after adjustment for clinicopathological characteristics. A nomogram was then constructed to estimate patient survival probability, supporting clinical decision-making. Expression of pyroptosis-related proteins was assessed using tissue proteomic data from the Clinical Proteomic Tumor Analysis Consortium (CPTAC) database. Additionally, the Tumor-Immune System Interaction Database (TISIDB) database was used to analyze immune cell infiltration, whereas gasdermin B (GSDMB) expression in ccRCC cell lines was assessed by Western blot (WB) and quantitative PCR (qPCR) experiments, with immunohistochemistry applied to measure the levels of GSDMB and programmed cell death protein 1 (PD-1).

A prognostic risk model was developed using nine genes related to pyroptosis ( AIM2 , CASP5 , GSDMB , GSDMD , NLRP7 , NOD2 , PYCARD , SCAF11 , and NLRP6 ), employing both univariate Cox and LASSO regression analyses. Based on the median risk score, ccRCC samples were divided into high- and low-risk categories, with Kaplan-Meier analysis revealing poorer survival rates for the high-risk group. Time-dependent receiver operating characteristic curve analysis confirmed the predictive performance of the model for ccRCC prognosis. Furthermore, a nomogram with calibration plots was developed to predict 1-, 3-, and 5-year survival probabilities for patients with ccRCC. Notably, these pyroptosis-related genes were strongly correlated with immune cell infiltration, including cytotoxic cells, CD56bright natural killer cells, and T helper cells. Subsequently, qPCR and WB analyses confirmed elevated GSDMB expression in ccRCC cells, and immunohistochemistry demonstrated a strong correlation between GSDMB and PD-1 expression.

Pyroptosis-associated genes, namely AIM2 , CASP5 , GSDMB , GSDMD , NLRP7 , NOD2 , PYCARD , SCAF11 , and NLRP6 , are promising prognostic biomarkers for ccRCC, and may serve as prognostic biomarkers for predicting patient survival and informing immunotherapy strategies.