Toll-like Receptor 6 and Connective Tissue Growth Factor are significantly upregulated in mitomycin-C treated urothelial carcinoma cells under hydrostatic pressure stimulation

Urothelial carcinoma (UC) is the most common histologic subtype of bladder cancer. The administration of mitomycin C (MMC) into bladder after transurethral resection of the bladder tumor (TURBT) is a common treatment strategy for preventing recurrence after surgery. We previously applied hydrostatic pressure combined with MMC in UC cells and found that hydrostatic pressure synergistically enhanced MMC-induced UC cell apoptosis via the Fas/FasL pathways. To understand the alteration of gene expressions in UC cells caused by hydrostatic pressure and MMC, oligonucleotide microarray was used to explore all of the differentially expressed genes. After bioinformatics analysis and gene annotation, Toll-like receptor 6 (TLR6) and connective tissue growth factor (CTGF) showed significant up-regulation among altered genes, and their expressions with each treatment of UC cells were validated by quantitative real-time PCR (qPCR). We conclude that under treatment with MMC and hydrostatic pressure, UC cells showed increasing apoptosis via extrinsic pathways through upregulation of TLR6 and CTGF. BFTC905 cells, a UC cell line, were cultured in the presence of 10 μg/ml of MMC for 2.5 hours. The MMC-treated cells were then incubated in a well-designed hydrostatic bioreactor for 10 kPa pressure treatment in duplicate or were cultured as usual to be used as control sets.