The role of IKK in constitutive activation of NF-kappa B transcription factor in prostate carcinoma cells

Rel/NF-kappaB transcription factors are implicated in the control of cell proliferation, apoptosis and transformation. The key to NF-kappaB regulation is the inhibitory IkappaB proteins. During response to diverse stimuli, IkappaBs are rapidly phosphorylated by IkappaB kinases (IKKs), ubiquitinated and undergo degradation. We have investigated the expression and function of NF-kappaB, IkappaB inhibitors and IKKs in normal prostate epithelial cells and prostate carcinoma (PC) cell lines LNCaP, MDA PCa 2b, DU145, PC3, and JCA1. We found that NF-kappaB was constitutively activated in human androgen- independent PC cell lines DU145, PC3, JCA1 as well as androgen- independent CL2 cells derived from LNCaP. In spite of a strong difference in constitutive kappaB binding, Western blot analysis did not reveal any significant variance in the expression of p50, p65, IkappaBs, IKKalpha, and IKKbeta between primary prostate cells, androgen-dependent and androgen- independent PC cells. However, we found that in androgen- independent PC cells IkappaBalpha was heavily phosphorylated and displayed a faster turnover. Using an in vitro kinase assay we demonstrated constitutive activation of IKK in androgen- independent PC cell lines. Blockage of NF-kappaB activity in PC cells by dominant-negative IkappaBalpha resulted in increased constitutive and TNF-alpha-induced apoptosis. Our data suggest that increased IKK activation leads to the constitutive activation of NF-kappaB 6 survival signaling' pathway in androgen-independent PC cells. This may be important for the support of their androgen-independent status and growth advantage.

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