Identification and functional characterization of EFNA3 as a dual diagnostic biomarker and immune therapeutic target in rectal cancer

Abstract

Background

The Eph/ephrin signaling system has emerged as an important regulator of oncogenic processes, yet its specific involvement in rectal adenocarcinoma (READ) pathogenesis requires further elucidation. This study employed an integrative analytical approach combining computational biology with experimental validation to characterize Eph/ephrin family members in READ, with emphasis on EFNA3.

Methods

We employed an integrative multi-omics approach combining bioinformatics analysis with experimental validation. Transcriptomic data from the Cancer Genome Atlas and Genotype-Tissue Expression were analyzed to evaluate Eph/ephrin family expression, clinical correlations, and immune infiltration patterns. Functional validation was performed using CCK-8, wound healing, transwell migration assays, and Western blotting.

Results

Our integrated multi-omics analysis identified EFNA3 as a dual-function biomarker with both prognostic and immunological relevance in READ. Specifically, EFNA3 expression was elevated in READ tissues and correlated with poor patient prognosis. Functional characterization revealed that EFNA3 contributes to an immunosuppressive tumor microenvironment, marked by reduced cytotoxic lymphocyte infiltration and downregulation of key immune checkpoints. Mechanistically, EFNA3 overexpression promoted rectal cancer cell proliferation and migration, with pathway analysis and Western blot validation implicating Phosphoinositide 3-Kinase (PI3K)/Ak strain Transforming (AKT)/Mechanistic Target of Rapamycin (mTOR) signaling activation. Furthermore, EFNA3 expression exhibited significant correlations with drug sensitivity and potential associations with traditional Chinese herbs, suggesting its broader implications in therapeutic response and alternative medicine approaches.

Conclusion

Our findings establish EFNA3 as a key regulator of READ progression, driving both tumor aggressiveness and immune evasion. These results provide a translational framework for targeting EFNA3-mediated pathways, suggesting combined PI3K/AKT/mTOR inhibition and immune modulation as a potential therapeutic strategy for aggressive READ.