Characterization of TGF-beta-regulated interleukin-8 expression in human prostate cancer cells

Prostate. 2006 Jun 15;66(9):996-1004. doi: 10.1002/pros.20424.

Abstract

Background: Interleukin (IL)-8 and transforming growth factor (TGF)-beta1 are overexpressed in advanced prostate cancer. The purpose of this study was to investigate TGF-beta1-regulated IL-8 expression in prostate cancer cells.

Methods: TGF-beta receptor expression was evaluated by real-time reverse-transcription PCR (RT-PCR) and Western blotting. TGF-beta1-regulated IL-8 expression was determined by real-time RT-PCR, enzyme-linked immunoabsorbance assay (ELISA), nuclear run-on, and IL-8 promoter reporter assay.

Results: PC-3MM2 cells expressed type I and type II TGF-beta receptors (TbetaRI and TbetaRII). LNCaP cells expressed significantly lower level of TbetaRII. Constitutive expression of IL-8 was detected in PC-3MM2 cells and LNCaP cells engineered with TbetaRII (LNCaP-TbetaRII). TGF-beta1 stimulated IL-8 expression in dose- and time-dependent manners, which was blocked by cycloheximide (CHX) and actinomycin D (ActD). The nuclear run-on and IL-8 luciferase reporter assays show that TGF-beta1 activated IL-8 gene transcription.

Conclusions: TGF-beta1 signaling regulates IL-8 expression in prostate cancer cells and may contribute to the overexpression of IL-8 in human prostate cancer.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Activin Receptors, Type I / analysis
  • Activin Receptors, Type I / genetics
  • Activin Receptors, Type I / physiology
  • Blotting, Western
  • Cell Line, Tumor
  • Cycloheximide / pharmacology
  • Dactinomycin / pharmacology
  • Dose-Response Relationship, Drug
  • Enzyme-Linked Immunosorbent Assay
  • Gene Expression Regulation, Neoplastic / drug effects*
  • Gene Expression Regulation, Neoplastic / physiology
  • Humans
  • Interleukin-8 / genetics*
  • Interleukin-8 / physiology
  • Male
  • Prostatic Neoplasms / genetics*
  • Prostatic Neoplasms / pathology
  • Protein Serine-Threonine Kinases
  • RNA, Messenger / analysis
  • RNA, Messenger / genetics
  • Receptor, Transforming Growth Factor-beta Type I
  • Receptor, Transforming Growth Factor-beta Type II
  • Receptors, Transforming Growth Factor beta / analysis
  • Receptors, Transforming Growth Factor beta / genetics
  • Receptors, Transforming Growth Factor beta / physiology
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction / physiology
  • Time Factors
  • Transcription, Genetic / drug effects
  • Transforming Growth Factor beta / genetics
  • Transforming Growth Factor beta / physiology*
  • Transforming Growth Factor beta1

Substances

  • Interleukin-8
  • RNA, Messenger
  • Receptors, Transforming Growth Factor beta
  • TGFB1 protein, human
  • Transforming Growth Factor beta
  • Transforming Growth Factor beta1
  • Dactinomycin
  • Cycloheximide
  • Protein Serine-Threonine Kinases
  • Activin Receptors, Type I
  • Receptor, Transforming Growth Factor-beta Type I
  • Receptor, Transforming Growth Factor-beta Type II