Potential Cross-Talk between Alternative and Classical NF-κB Pathways in Prostate Cancer Tissues as Measured by a Multi-Staining Immunofluorescence Co-Localization Assay

PLoS One. 2015 Jul 17;10(7):e0131024. doi: 10.1371/journal.pone.0131024. eCollection 2015.

Abstract

Background: While the classical NF-κB/p65 pathway is known to be involved in prostate cancer progression and is associated with poor patient outcome, the role of the NF-κB /RelB alternative protein is not well defined. Here we analyzed the activation of both NF-κB pathways in prostate cancer tissues and correlate this activation with clinical features of the disease.

Methods: A multiple immunofluorescence technique was employed to concomitantly and quantitatively visualize the nuclear localization of p65 and RelB in 200 paraffin embedded samples. Epithelia were defined using appropriate fluorochrome markers and the resulting immunofluorescent signals were quantified with an automated scoring system.

Results: The nuclear frequency of p65 was found to be significantly increased in tumor tissues as compared with normal adjacent tissue, whereas the frequency for RelB was decreased (p < 0.001, Wilcoxon test). As previously reported, p65 nuclear frequency was associated with a risk of biochemical recurrence. Although, RelB nuclear frequency alone did not predict recurrence, the presence of activated RelB reduced the risk of recurrence associated with the activation of p65.

Conclusion: For the first time p65/RelB co-distribution was assessed in prostate cancer tissues and suggested a negative crosstalk between the two NF-κB pathways in prostate cancer progression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Nucleus / metabolism*
  • Cell Nucleus / ultrastructure
  • Cohort Studies
  • Disease Progression
  • Fluorescent Antibody Technique
  • Gene Expression Regulation, Neoplastic*
  • Humans
  • Male
  • Microtomy
  • Neoplasm Recurrence, Local / diagnosis
  • Neoplasm Recurrence, Local / metabolism*
  • Neoplasm Recurrence, Local / pathology
  • Neoplasm Recurrence, Local / surgery
  • Paraffin Embedding
  • Prognosis
  • Prostatectomy
  • Prostatic Neoplasms / diagnosis
  • Prostatic Neoplasms / metabolism*
  • Prostatic Neoplasms / pathology
  • Prostatic Neoplasms / surgery
  • Signal Transduction
  • Staining and Labeling
  • Tissue Array Analysis
  • Transcription Factor RelA / genetics
  • Transcription Factor RelA / metabolism*
  • Transcription Factor RelB / genetics
  • Transcription Factor RelB / metabolism*

Substances

  • RELB protein, human
  • Transcription Factor RelA
  • Transcription Factor RelB

Grants and funding

Chair en Cancer de la prostate (Université de montreal, http://www.recherche.umontreal.ca/en/research-at-udem/our-research-units/profile/unite/449/pid/15/) CUOG award (http://www.cuog.ca) Réseau de recheche en cancer (http://www.rrcancer.ca/fr/scientifique/biobanques/lister/2). Visiopharm' provided support in the form of a salary for author TK, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific role of this author is articulated in the ‘author contributions’ section.