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

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Annual rept. 1 Aug 2004-31 Jul 2005

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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-KB prostate carcinoma PC cells. During FYO2 we developed the conditions for RNA isolation from OCT-embedded frozen PC and BPH samples, developed conditions for cell lysis and IKKi immunoprecipitation from transfected cells using FLAG antibody. We also generated PC cell lines stably infected with lentiviruses harboring w.t. IKKi, d.n. IKKi mutant K38A, and constitutively active IKKi mutant S172E. We continued the characterization of biological effects of IKKi overexpressionIKKi blockage in PC cells. Our data provide the experimental evidence that IKKi could be involved in the regulation of NF-kB activity in PC cells through a positive feedback loop. Inactivation of IKKi in PC cells via infection with lentivirus expressing IKKi d.n. mutant indicated that IKKi plays an important role in the regulation not only NF-kB but also other transcriptional factors. IKKi appeared to be very important for PC cell growth regulation. The in vitro growth of PC3 cells infected with IKKi d.n. mutant was reduced by 60. The results of our studies have been presented at the local and national meetings, two manuscripts are published, one is in press, and the fourth manuscript is under preparation.

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  • Medicine and Medical Research
  • Statistics and Probability

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