Comprehensive analysis of ALYREF gene expression and its correlation with immunotherapy efficacy and circulating tumor cells in bladder cancer

Bladder cancer (BCa) presents a significant clinical challenge, with a pressing need for biomarkers to predict prognosis and guide immunotherapy. Aly/REF export factor (ALYREF) is a key regulator in various cancers, but its role in bladder cancer (BCa) remains unclear. This study aimed to investigate the clinical significance of ALYREF and its association with immunotherapy response and metastatic potential in BCa. We integrated pan-cancer bioinformatic analyses with clinical validation using RT-qPCR, immunohistochemistry, and Western blot on patient samples. We assessed ALYREF's correlation with clinicopathological features, survival, and response to immune checkpoint inhibitors (ICIs). Circulating tumor cell (CTC) counts were analyzed to link ALYREF to metastatic potential. Functional roles in migration and invasion were validated in vitro using the T24 cell line. ALYREF was significantly upregulated in BCa tissues, correlating with higher tumor grade and poorer overall survival. Paradoxically, high ALYREF expression was also associated with a better response to ICI therapy. Furthermore, elevated ALYREF levels in tumors corresponded to increased CTC counts. In vitro experiments confirmed that ALYREF promotes BCa cell migration and invasion. ALYREF is a dual-role biomarker in BCa. Its overexpression signifies aggressive tumor biology and metastatic risk yet also predicts a favorable response to immunotherapy. ALYREF holds promise for refining patient stratification and personalizing BCa treatment.