Constitutive Activation of NF-kappaB in Prostate Carcinoma Cells Through a Positive Feedback Loop: Implication of Inducible IKK-Related Kinase (IKKi)

摘要

The overall goal of this project is to understand the role of inducible IKK-related kinase IKKi in constitutive activation of anti-apoptotic transcription factor NF-kappaB prostate carcinoma (PC) cells. During FY03 we developed the optimal conditions for prostate cell PC cell stable infection by lentiviruses; regimens for selection of infected cells; and enrichment of cells co-infected with yellow fluorescent protein (YFP) by FACS. We also developed a test for monitoring of growth of live cell cultures infected with YFP using fluorescent plate reader. We continued the development of the conditions for IKKi immunoprecipitation to reveal the content of IKKi native complexes in PC cells. We assessed the biological effect of IKKi on prostate carcinoma cells. We found that IKKi in LNCaP prostate cells significantly increased their growth in vitro, increased their tumorigenicity. Our experiments indicated that IKKi affected the transcriptional activity of androgen receptor. These results suggest an important role of IKKi in the transition of prostate cells to androgen-independent growth. The results of our studies have been presented at the local and national meetings, one manuscript has been published, one is in press, one is under revision, and fourth is under preparation.