Cigarette smoke supports stemness and epithelial-mesenchymal transition in bladder cancer stem cells through SHH signaling

Cancer stem cells (CSCs) are essential in every step of tumorigenesis and progression. As an important process in cancer development, epithelial-mesenchymal transition (EMT) has been reported to promote stem-like cells. Bladder cancer is one of the most common cancers in the urinary tract, and cigarette smoke (CS) is a preventable risk factor. In the present study, we tested the hypothesis that CS could promote stemness and EMT in bladder cancer. Bladder cancer UM-UC-3 and EJ cell lines were maintained in serum-free medium to grow as tumor spheres, characteristic of CSCs. Results demonstrated that CS enhanced tumor sphere formation capacity, upregulated expression of CSC markers, increased the proportion of the CD44⁺ cell population, and promoted EMT. Mechanistically, the Sonic Hedgehog (SHH) pathway regulated CS-triggered EMT and stemness. More importantly, among bladder cancer patients, smokers harbored higher levels of CSC markers and proteins for SHH signaling than non-smokers. Collectively, findings in this study highlight the critical role of CS in the stemness and EMT of bladder cancer. Smoking cessation and intervening in the SHH pathway may both be strategies to prevent bladder cancer.