The result of differential expression analysis.

BackgroundLymph node metastasis (LNM) is a key prognostic factor in colorectal cancer (CRC), and early lymph node status assessment is crucial for prognosis and immunotherapy decisions. However, the immune characteristics of LNM-negative CRC remain poorly understood.MethodsUsing machine learning algorithms, we identified and analyzed two immune subtypes (C1 and C2) in 244 LNM-negative CRC samples, with validation in 458 additional samples, and evaluated their immune characteristics and functional pathways.ResultsSubtype C1 exhibited high immune scores and responsiveness to immune checkpoint blockade, while subtype C2 showed a favorable prognosis and increased immune cell infiltration, indicating it represents an earlier CRC stage. Experimental validation revealed that PPFIA4 knockdown in C1 significantly suppressed CRC cell proliferation and migration.ConclusionsThese findings provide insights into personalized immunotherapy strategies for early-stage CRC patients and have potential clinical application value.