Cytokine-driven modulation of WT1 and IL-10 in lung cancer progression

Lung cancer is driven by complex interactions between oncogenes and the inflammatory microenvironment. In particular, the cytokines tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β), and their modulation of key regulators like Wilms' tumor 1 (WT1) and interleukin-10 (IL-10) remain underexplored. This study aims to investigate the role of these cytokines in WT1 and IL-10 regulation during lung cancer progression.

A total of 982 lung cancer patient samples from The Human Protein Atlas were analyzed. In vitro, RAW264.7 macrophages were transfected with a WT1 plasmid (pWT1) and treated with TNF-α, IL-1β, and lipopolysaccharide (LPS). WT1 and IL-10 expression was evaluated in A549, B16-F10, and J774.2 cell lines using reverse transcription polymerase chain reaction (RT-PCR), Western blotting, and enzyme-linked immunosorbent assay (ELISA). Immunofluorescence was employed to assess WT1 localization and phosphorylation, while immunohistochemistry was used to evaluate the correlation between WT1 and IL-10 in patient samples.

WT1 expression progressively increased from stage I to IV lung cancer and positively correlated with IL-10 in stages II and IV. WT1 overexpression in RAW264.7 cells treated with LPS led to a 12.9-fold increase in IL-10 expression. Proinflammatory cytokines decreased WT1 in A549 and B16-F10 cells but increased it in J774.2 macrophages, leading to cytoplasmic localization and phosphorylation. Patient sample analysis revealed a positive correlation between WT1 and IL-10 in advanced stages.

These findings suggest that WT1 and IL-10 are modulated by inflammatory cytokines in a stage-dependent manner in lung cancer. WT1 upregulation is associated with increased IL-10 expression, particularly in advanced stages, highlighting potential therapeutic targets for modulating the immune response in lung cancer.