CFTR and SLC26A6 have been previously demonstrated to be expressed in the male and female reproductive systems and play an important role in pH regulation. However their expression and function in the prostate are unclear. To investigate this, the present study established a primary culture of rat ventral prostate epithelium grown on permeable supports and the expression of CFTR and SLC26A6 was determined by RT-PCR, western blot and immunostaining. The results obtained by these methods consistently showed the expression of CFTR and SLC26A6 in rat prostate epithelial cells. Experiments were also carried out to demonstrate the involvement of CFTR and SLC26A6 in pH regulation across the cultured prostate epithelial cells by fluorimetric intracellular pH measurements using the pH-sensitive fluorescent dye BCECF-AM. The results showed that the pH recovery from alkalization could be altered by forskolin (10µM) and NPPB (100µM), agents known to active and inhibit CFTR, respectively. The physiological function of both CFTR and SLC26A6 in the rat prostate epithelium and their possible involvement in the pathogenesis of prostate diseases await further studies.