Previous studies have shown that the hamster CYP11B2 gene promoter is under the influence of angiotensin II (AII), cAMP and potassium (K+). However, very little is known about the mechanisms by which these compounds regulate the transcription of the CYP11B2 gene. Therefore we analysed the 5′-flanking region of the hamster CYP11B2 gene using a transient transfection expression system in NCI-H295 adrenocortical cells, which are known to respond to K+, cAMP and AII. The first 486 bp before the transcription initiation site were introduced upstream of the chloramphenicol acetyl transferase gene. NCI-H295 cells transfected with this - 486 construct showed increased CAT activity upon treatment by K+, All, forskolin and cAMP. The calcium channel antagonist nifedipine partially blocked the enhancing effects of All, forskolin and cAMP by 35%, 30% and 30% respectively, whereas it completely blocked the stimulatory effects of KC1 (1). These results thus show the involvement of calcium channels in the regulation of CYP11B2 gene transcription by K+, and their partial involvement in the regulation of this gene by AII, forskolin and cAMP in NCI-H295 cells.