Phases of tight junction barrier disruption during transurothelial migration of invasive urothelial cancer cells

Bladder cancer is characterised by its multifocal nature and a high recurrence, yet the underlying mechanisms of these phenomena remain only partially understood. In the present study, we aimed to investigate transurothelial invasion of urothelial cancer cells as a potential mechanism for dissemination of bladder cancer and to identify the key molecules involved in urothelial barrier disruption. Using confocal and electron microscopy, we were able to show that within a 24-hour timeframe muscle-invasive urothelial cancer cells T24 adhere to the partially differentiated normal urothelial in vitro model and initially cause localised disruption of the tight junctions between urothelial cells. Subsequently, urothelial cells separate and individual T24 cells migrate paracellularly through the urothelium. qPCR analysis identified fibroblast activation protein (FAP)/seprase as the candidate most likely to be involved in urothelial barrier disruption. In addition, treatment of T24 cells with Pefabloc resulted in the inhibition of T24 cell invasion. Our results contribute to the understanding of the mechanisms underlying transurothelial invasion of urothelial cancer cells. Among the molecules tested, FAP/sepraseis likely involved in cancer cell-induced disruption of the urothelial barrier, suggesting its potential as a therapeutic target to prevent progression and recurrence of bladder cancer.