Background: Pyroptosis, an inflammatory form of programmed cell death, has been implicated in tumor progression, yet its molecular contribution across breast cancer subtypes remains poorly defined. Methods: To characterize pyroptosis-related alterations, we analyzed tumor and matched control tissues from five molecular subtypes of breast cancer using genome-wide messenger RNA and microRNA microarrays, quantitative polymerase chain reaction, enzyme-linked immunosorbent assays, and protein–protein interaction analysis. We identified predicted microRNA–messenger RNA regulatory relationships and constructed a pyroptosis index and inflammasome activation score. To contextualize these findings, temporal expression changes were evaluated in a cryoablation model of benign fibroadenoma. Results: Nine genes associated with inflammatory and apoptotic signaling— CXCL8 , BCL2 , BAX , CASP1 , CASP9 , TP53 , CDKN1A , CDKN1B , and MMP9 —consistently distinguished cancerous from control tissue across all subtypes at both messenger RNA and protein levels. Aggressive subtypes, particularly human epidermal growth factor receptor 2–enriched and triple-negative tumors, exhibited pronounced activation of inflammasome-related pathways, elevated pyroptosis index and inflammasome activation score values, and coordinated suppression of cell-cycle inhibitors. Predicted microRNA regulators, including microRNA 140-3p, microRNA 124-3p, microRNA 300, microRNA 30a-3p, microRNA 30d-3p, and microRNA 608, showed patterns consistent with loss of post-transcriptional restraint in high-grade tumors. In fibroadenoma, pyroptosis-associated expression changes were rapid and transient, whereas malignant tissue displayed a consistent, subtype-dependent elevation of pyroptosis-related markers at the time of resection. Conclusion: This integrative analysis identifies a conserved pyroptosis-related molecular signature that deepens understanding of inflammatory programmed cell death in breast cancer and highlights interconnected pathways with diagnostic, prognostic, and therapeutic relevance.

Apoptosis maintains tissue homeostasis, and its dysregulation is closely associated with breast cancer progression and therapeutic resistance. We performed an integrative analysis of apoptosis-related signaling in breast cancer tissues across five molecular subtypes and compared these patterns with systemic apoptotic responses following cryoablation of benign fibroadenomas. Gene expression profiling was conducted using mRNA microarrays and validated by qRT-PCR and ELISA. Apoptosis pathway activity was assessed with the MSigDB HALLMARK_APOPTOSIS gene set, including intrinsic and extrinsic pathway scoring and an apoptotic balance index (ABI). MicroRNA profiling combined with in silico analyses identified potential miRNA–mRNA interactions. A progressive shift toward reduced pro-apoptotic and enhanced stress-adaptive signaling was observed with increasing tumor aggressiveness, most pronounced in triple-negative and non-luminal HER2-positive cancers. This pattern included reduced intrinsic pathway activity, decreased ABI, downregulation of pro-apoptotic genes (BIK, BMF, TXNIP), and upregulation of stress-associated or cytoprotective genes (HSPB1, PPT1). Several expression patterns were accompanied by overexpression of miRNAs (miR-582-5p, miR-421, miR-106b-5p, miR-20a-5p, miR-20b-5p, miR-93-5p) predicted to target apoptosis-related genes. In contrast, fibroadenoma cryoablation was associated with transient systemic modulation of apoptosis-related genes and proteins followed by gradual normalization. These findings highlight differences between apoptosis-related dysregulation in malignant tissue and regulated systemic responses following benign tissue injury, supporting pathway-level interpretation and identifying candidate molecular networks warranting further mechanistic and translational investigation.

Ferroptosis is an iron-dependent form of regulated cell death driven by lipid peroxidation and oxidative stress, increasingly implicated in cancer biology. However, its molecular regulation across breast cancer subtypes and its potential systemic manifestations remain incompletely understood. The aim of this study was to identify ferroptosis-associated molecular alterations that are largely shared across subtypes and to evaluate their systemic reflection following localized tissue injury. Tumor and matched normal breast tissues representing major molecular subtypes were analyzed. Global mRNA and miRNA expression profiling was performed using microarrays, followed by validation of selected genes using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). Functional enrichment and protein–protein interaction analyses were conducted to characterize associated pathways. In addition, systemic responses were assessed in patients undergoing fibroadenoma cryoablation through longitudinal blood sampling. Six ferroptosis-related genes (SLC7A11, GPX4, FTH1, NQO1, NFE2L2, SQSTM1) demonstrated consistent upregulation across all breast cancer subtypes, with higher expression observed in more aggressive tumors. These genes are functionally linked to antioxidant defense, iron metabolism, and oxidative stress regulation, and their coordinated expression pattern is consistent with activation of NRF2-dependent cytoprotective pathways. Downregulation of selected miRNAs may contribute to this expression profile but likely represents a secondary regulatory mechanism. Survival analysis revealed heterogeneous and subtype-dependent associations, with limited and gene-specific prognostic relevance. Cryoablation induced transient increases in circulating levels of the analyzed proteins, reflecting systemic responses to localized tissue injury. In conclusion, breast cancer is characterized by a largely shared ferroptosis-associated molecular signature across subtypes; however, its clinical impact appears to be variable and context-dependent. Systemic detection of related molecular signals suggests potential utility as indicators of tissue stress responses, although their role as specific biomarkers of ferroptosis requires further validation.