Ellagic Acid Resensitizes Gemcitabine-Resistant Bladder Cancer Cells by Inhibiting Epithelial-Mesenchymal Transition and Gemcitabine Transporters

SIMPLE SUMMARY: Chemoresistance of bladder cancer has become a major obstacle to clinical treatment, especially in first-line treatments involving gemcitabine (GCB). Epithelial-mesenchymal transition (EMT) is highly correlated with GCB resistance but less correlated with GCB metabolism and less reported as a novel therapeutic strategy. Our findings indicated that EMT-related GCB resistance occurs through the TGF-β/Smad signaling pathways and involves repressed expression of the GCB transporters hCNT1 and hENT1. Ellagic acid (EA) combined with GCB intensified the chemosensitivity of GCB in resistant cells by repressing Smad2, Smad3, and Smad4 expression and rescuing hCNT1 and hENT transcription. These data suggest that EA is a good adjuvant agent for blocking TGF-β/Smad signaling-related GCB resistance in bladder cancer. ABSTRACT: Gemcitabine (GCB) resistance is a major issue in bladder cancer chemoresistance, but its underlying mechanism has not been determined. Epithelial-mesenchymal transition (EMT) has been shown to be comprehensively involved in GCB resistance in several other cancer types, but the direct connection between EMT and GCB remains unclear. This study was designed to elucidate the mechanism of EMT-related GCB resistance in bladder cancer and identify a potential phytochemical to modulate drug sensitivity. The biological effects of ellagic acid (EA) or its combined effects with GCB were compared in GCB-resistant cells and the GCB-sensitive line in terms of cell viability, apoptosis, motility, and in vivo tumorigenicity. The molecular regulation of EMT-related GCB resistance was evaluated at both the mRNA and protein expression levels. Our results indicated that TGF-β/Smad induced the overactivation of EMT in GCB-resistant cells and reduced the expression of GCB influx transporters (hCNT1 and hENT1). Moreover, ellagic acid (EA) inhibited the TGF-β signaling pathway both in vitro and in vivo by reducing Smad2, Smad3, and Smad4 expression and thereby resensitized GCB sensitivity. In conclusion, our results demonstrate that TGF-β/Smad-induced EMT contributes to GCB resistance in bladder cancer by reducing GCB influx and also elucidate the novel mechanisms of EA-mediated inhibition of TGF-β/Smad-induced EMT to overcome GCB resistance. Our study warrants further investigation of EA as an effective therapeutic adjuvant agent for overcoming GCB resistance in bladder cancer.