Androgens regulate protein kinase Cdelta transcription and modulate its apoptotic function in prostate cancer cells

Cancer Res. 2006 Dec 15;66(24):11792-801. doi: 10.1158/0008-5472.CAN-06-1139.

Abstract

Activation of protein kinase Cdelta (PKCdelta), a member of the novel PKC family, leads to apoptosis in several cell types. Although the molecular bases of PKCdelta activation are being unfolded, limited information is available on the mechanisms that control its expression. Here, we report that in prostate cancer cells PKCdelta is tightly regulated by androgens at the transcriptional level. Steroid depletion from the culture medium causes a pronounced down-regulation of PKCdelta protein and mRNA in androgen-sensitive LNCaP prostate cancer cells, an effect that is rescued by the androgen R1881 in an androgen receptor (AR)-dependent manner. Analysis of the PKCdelta promoter revealed a putative androgen responsive element (ARE) located 4.7 kb upstream from the transcription start site. Luciferase reporter assays show that this element is highly responsive to androgens, and mutations in key nucleotides in the AR-binding consensus abolish reporter activity. Furthermore, using chromatin immunoprecipitation assays, we determined that the AR binds in vivo to the PKCdelta ARE in response to androgen stimulation. Functional studies revealed that, notably, androgens modulate phorbol 12-myristate 13-acetate (PMA)-induced apoptosis in LNCaP cells, an effect that is dependent on PKCdelta. Indeed, androgen depletion or AR RNA interference severely impaired the apoptotic function of PKCdelta or the activation of p38, a downstream effector of PKCdelta in LNCaP cells--effects that can be rescued by restoring PKCdelta levels using an adenoviral delivery approach. Our studies identified a novel hormonal mechanism for the control of PKCdelta expression via transcriptional regulation that fine-tunes the magnitude of PKCdelta apoptotic responses.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Apoptosis / drug effects*
  • Base Sequence
  • Cell Line, Tumor
  • DNA Primers
  • Gene Expression Regulation, Enzymologic / drug effects
  • Gene Expression Regulation, Neoplastic / drug effects
  • Genes, Reporter
  • Humans
  • Isoenzymes / genetics
  • Isoenzymes / metabolism
  • Male
  • Metribolone / pharmacology*
  • Promoter Regions, Genetic
  • Prostatic Neoplasms / enzymology
  • Prostatic Neoplasms / genetics*
  • Prostatic Neoplasms / pathology
  • Protein Kinase C-delta / genetics*
  • Protein Kinase C-delta / metabolism
  • RNA Interference
  • Restriction Mapping
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcription, Genetic / drug effects*
  • Transfection

Substances

  • DNA Primers
  • Isoenzymes
  • Metribolone
  • Protein Kinase C-delta