Role of IKKs and Transcription Factor NF-kB in Prostate Tumorigenesis
Abstract
The overall goal of this project is to understand the role of up-stream IkB kinases and NF-xB "survival signaling" pathway in tumorigenesis in prostate. We found previously that NF-kB was constitutively activated in human androgen-independent prostate carcinoma (PC) cell lines due to the activation of IKKalpha/beta kinases. We have generated clones of PC cells transfected with IKK d.n. mutants but in most of those clones the expression of IKKbeta d.n. and IKKalpha d.n. mutants significantly decreased during cell cultivation. To overcome the transgene silencing, we have undertaken an alternative approach and performed the experiments with highly specific chemical inhibitor of IKKbeta, PS1145. We found that PS1145 inhibited basal and inducible NF-kB in PC cells, sensitized PC cells to apoptosis, and reduced their metastatic potential. We have also started to generate inducible IKKalpha d.n. and IKKbeta d.n. constructs using retroviral Tet-On expression system (Clontech Laboratories). We also studied the expression of IKKs and NF-kB in prostate tissues. The analysis of immunostaining of NF-kB and IKK proteins in BPH (benign prostate hyperplasia) and PC samples has revealed modest increase of p65 expression in advanced PCs. There were no significant changes in the expression of p50 and IKKbeta in PC in comparison to BPH. P52 expression was significantly higher in high grade PCs in comparison to BPH and low grade PCs. Importantly we found frequent nuclear localization of p52 in PCs. In some tumors p52 immunostaining pattern correlated with IKKalpha expression, and especially with the level of IKKalpha/beta phosphorylation.
Document Details
- Document Type
- Technical Report
- Publication Date
- May 01, 2004
- Accession Number
- ADA427879
Entities
People
- Irina Budunova
Organizations
- Northwestern University