ANDROGEN REGULATION OF PROSTATE GROWTH

  • Tindall, Donald J (PI)

Project: Research project

Project Details

Description

Prostate cells require the presence of androgens for differentiation,
development and maintenance. The effects of androgens are mediated
predominantly by the androgen receptor (AR). Recent studies have
demonstrated that expression of the AR gene is regulated by androgens and
modulated by other growth regulatory factors. These studies suggest that
both androgens and other growth regulatory factors (i.e. retinoic acid
(RA), TPA via c-Jun/c-Fos and the cAMP regulated protein, CREB) play
roles in regulating the AR gene activity. In order to define these roles
the following specific aims will be addressed: Specific Aim 1 proposes
to determine whether androgens and other factors regulate the synthesis
and/or stabilization of AR mRNA and protein by a) Nuclear run-on analysis
of AR and mRNA in human prostate cells treated with androgen plus/minus
RA, TPA and forskolin, b) Measurement of the half-life of AR mRNA in the
presence of androgen plus/minus RA, TPA and forskolin c) Northern blot
analysis of AR, CREB, c-Jun and c-Fos mRNAs in cells treated with
androgen plus/minus RA, TPA and forskolin. Specific Aim 2 proposes to
identify transcriptional regulatory elements in the promoter region of
the human AR gene by a) Cloning and sequencing the 5'-flanking region of
the human androgen receptor gene, b) Cotransfection of the hAR promoter
(full-length and deletion mutants) into prostate cells with vectors
expressing hAR, RAR, cJun, cFos and CREB and c) Footprinting analysis of
protein binding sites in the promoter region of the hAR gene in prostate
cells transfected with hAR, RAR, cJun, cFos, and CREB. Specific Aim 3
proposes to characterize the mechanism(s) by which transcription factors
regulate the human AR gene by a) Cotransfection of hAR, RAR, cJun, cFos
and CREB with putative regulatory elements (wild-type and mutated)
coupled to a heterologous promoter (tk-CAT) and b) Band-shift analysis
of putative regulatory elements with (i) nuclear extracts from prostate
cells treated with androgen plus/minus RA, TPA and forskolin and (ii)
recombinant nuclear transcription factors (AR, RAR, cJun, cFos, and
CREB).
StatusFinished
Effective start/end date9/30/938/31/97

Funding

  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health

ASJC

  • Medicine(all)

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