Cancer-associated fibroblasts from cervical cancer modulate protumoral neutrophil functions: evidence from a pilot exploratory study

Cancer-associated fibroblasts (CAFs) are key regulators of the tumor microenvironment; however, their role in modulating neutrophil function during cervical cancer (CC) progression remains poorly understood. In this study, we evaluated whether CAF supernatants (CSN) derived from early (FIGO I-II) and late stages (FIGO III-IV) of CC differentially regulate neutrophil activity. Peripheral blood neutrophils from healthy donors were stimulated with CSN, and their functional responses were assessed, including cytokine production, metabolic activity (MTT), reactive oxygen species (ROS) generation, apoptosis and survival analyses, expression of activation (CD66b) and protease degranulation markers (MMP9 and NE), and neutrophil extracellular trap (NET) release. CSN factors promoted a predominantly proinflammatory cytokine profile in neutrophils, characterized by increased production of IL-6, IL-8, TNF-α, and IFN-γ. Additionally, stimulation with CAF enhanced neutrophil metabolic activity, ROS production, survival, and expression of CD66b, MMP9, and neutrophil elastase, suggesting preliminary stage-associated differences in neutrophils responses between CAFs derived from early- and late-stage tumors. Notably, NET formation was significantly higher in neutrophils stimulated with early-stage CAFs, whereas late-stage CAFs induced a metabolically active but lower NETosis release. These exploratory findings suggest that cervical cancer-derived CAFs may differentially modulate neutrophil functional responses according to tumor stage, highlighting the CAF-neutrophil axis as a potentially relevant component of the cervical cancer microenvironment.