Increased androgen receptor (AR) levels are associated with prostate cancer progression to androgen independence and therapy resistance. Evidence has suggested that chronic inflammation is closely linked to various cancers including prostate cancer. Herein we show that the pro-inflammatory cytokine TNF negatively regulates AR mRNA and protein expression, and reduces androgen sensitivity in androgen-dependent LNCaP human prostate cancer cells. Decreased AR expression results from transcription repression involving essential in cis interaction of NF-B with the B-myb transcription factor at a composite genomic element in the 5' UTR of AR. The negative regulation was abrogated when NF-B activity was inhibited by a superrepressor of the inhibitory kappaB (IB) protein. Contrastingly, androgen-independent C4-2 (LNCaP-derived) cells fail to show AR down regulation by TNF, despite expression of B-myb and TNF-induced NF-B activity similarly to that in LNCaP cells. The negatively regulated AR gene chromatin region showed TNF-dependent enrichment of B-myb and the NF-B proteins p65 and p50. In parallel, the histone deacetylase HDAC1, corepressor SMRT and the corepressor-associated scaffold protein mSin3A were recruited to the inhibitory site. In C4-2 cells, neither NF-B and B-myb, nor any of the corepressor components, were detected at the negative site in response to TNF. Apoptosis was induced in TNF-treated LNCaP cells, likely in part due to the down regulation of AR. The androgen-independent, AR-expressing C4-2 and C4-2B (derived from C4-2) cells were resistant to TNF-induced apoptosis. The results linking androgen dependence to NF-B and AR pathways may be insightful in identifying novel treatment targets for prostate cancer.