Skip to main content
  • AACR Journals
    • Blood Cancer Discovery
    • Cancer Discovery
    • Cancer Epidemiology, Biomarkers & Prevention
    • Cancer Immunology Research
    • Cancer Prevention Research
    • Cancer Research
    • Clinical Cancer Research
    • Molecular Cancer Research
    • Molecular Cancer Therapeutics

  • Register
  • Log in
  • My Cart
Advertisement

Main menu

  • Home
  • About
    • The Journal
    • AACR Journals
    • Subscriptions
    • Permissions and Reprints
    • Reviewing
  • Articles
    • OnlineFirst
    • Current Issue
    • Past Issues
    • Meeting Abstracts
    • Collections
      • Focus on Radiation Oncology
      • Novel Combinations
      • Reviews
      • Editors' Picks
  • For Authors
    • Information for Authors
    • Author Services
    • Best of: Author Profiles
    • Submit
  • Alerts
    • Table of Contents
    • OnlineFirst
    • Editors' Picks
    • Citation
    • Author/Keyword
  • News
    • Cancer Discovery News
  • AACR Journals
    • Blood Cancer Discovery
    • Cancer Discovery
    • Cancer Epidemiology, Biomarkers & Prevention
    • Cancer Immunology Research
    • Cancer Prevention Research
    • Cancer Research
    • Clinical Cancer Research
    • Molecular Cancer Research
    • Molecular Cancer Therapeutics

User menu

  • Register
  • Log in
  • My Cart

Search

  • Advanced search
Molecular Cancer Therapeutics
Molecular Cancer Therapeutics

Advanced Search

  • Home
  • About
    • The Journal
    • AACR Journals
    • Subscriptions
    • Permissions and Reprints
    • Reviewing
  • Articles
    • OnlineFirst
    • Current Issue
    • Past Issues
    • Meeting Abstracts
    • Collections
      • Focus on Radiation Oncology
      • Novel Combinations
      • Reviews
      • Editors' Picks
  • For Authors
    • Information for Authors
    • Author Services
    • Best of: Author Profiles
    • Submit
  • Alerts
    • Table of Contents
    • OnlineFirst
    • Editors' Picks
    • Citation
    • Author/Keyword
  • News
    • Cancer Discovery News
Research Articles: Therapeutics, Targets, and Development

Suberoylanilide hydroxamic acid (vorinostat) represses androgen receptor expression and acts synergistically with an androgen receptor antagonist to inhibit prostate cancer cell proliferation

Deborah L. Marrocco, Wayne D. Tilley, Tina Bianco-Miotto, Andreas Evdokiou, Howard I. Scher, Richard A. Rifkind, Paul A. Marks, Victoria M. Richon and Lisa M. Butler
Deborah L. Marrocco
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Wayne D. Tilley
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Tina Bianco-Miotto
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Andreas Evdokiou
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Howard I. Scher
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Richard A. Rifkind
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Paul A. Marks
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Victoria M. Richon
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Lisa M. Butler
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
DOI: 10.1158/1535-7163.MCT-06-0144 Published January 2007
  • Article
  • Figures & Data
  • Info & Metrics
  • PDF
Loading

Article Figures & Data

Figures

  • Figure 1.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 1.

    Inhibition of LNCaP and PC-3 prostate cancer cell growth by SAHA. LNCaP and PC-3 cells (2.5 × 104 per well in 24-well plates) were cultured with SAHA (0, 0.5, 1, 2.5, 5, 7.5, or 10 μmol/L) in RPMI 1640 containing 10% FCS, for up to 7 d. Cells were counted every day (PC-3) or every second day (LNCaP) using a hemocytometer, and cell viability was assessed by trypan blue dye exclusion (A and C). The number of dead cells is expressed as a percentage of total cells counted (B and D). Representative of at least three independent experiments. Points, mean of triplicate wells; bars, SE.

  • Figure 2.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 2.

    Effect of SAHA on cell cycle distribution in LNCaP cells. A, cells cultured with SAHA (0, 2.5, or 7.5 μmol/L) for 24 h were harvested by trypsinization, fixed in 70% ethanol, and stained with propidium iodide. Cell cycle distribution was determined by flow cytometry. B, cells cultured in the absence or presence of SAHA (7.5 μmol/L) for 24 h or 4 d were harvested and fixed as described above and stained with propidium iodide. The fraction of hypodiploid (sub-G1) nuclei was measured by flow cytometry using standard histogram analysis.

  • Figure 3.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 3.

    Induction of LNCaP cell death by SAHA. A, LNCaP cells (2.5 × 104 per well in 24-well plates) were cultured with SAHA (0, 2.5, or 7.5 μmol/L) with or without the z-VAD-fmk caspase inhibitor for 48 h and assayed for caspase-3 activity. B, LNCaP cells (2.5 × 104) were cultured with SAHA (0, 2.5, or 7.5 μmol/L) with or without the z-VAD-fmk caspase inhibitor for 5 d. Cells were counted at day 5 using a hemocytometer, and cell viability was assessed by trypan blue exclusion. The number of dead cells expressed as a percentage of total cells was counted. Columns, mean of triplicate wells in a representative experiment; bars, SE. C, DAPI staining of cell nuclei. LNCaP cells were seeded onto plastic coverslips and cultured with vehicle alone (DMSO) or SAHA (7.5 μmol/L) for 24 h. Cells were fixed with methanol and incubated with DAPI, before washing in PBS and mounting on PBS/glycerin. DAPI staining was visualized by fluorescence microscopy. D, effect of SAHA on mitochondrial membrane potential in LNCaP cells. Cells were cultured with vehicle alone (DMSO) or SAHA (7.5 μmol/L) for 48 h. Cells were harvested by trypsinization, resuspended in RPMI 1640 containing 10% FCS, and incubated with the mitochondrial dye rhodamine 123 (Rho123; 2 μg/mL) for 20 min at 37°C. Cells were washed and incubated in PBS containing 2 μg/mL of the viability dye 7-aminoactinomycin D at room temperature for 10 min and then analyzed by flow cytometry. Results shown indicate the level of rhodamine 123 fluorescence in the 7-aminoactinomycin D–negative population.

  • Figure 4.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 4.

    Summary of gene expression changes in LNCaP cells following SAHA treatment. A, scatter plot of control intensity versus SAHA (2.5 μmol/L) intensity for all elements on the UniGEM version 2.0 human cDNA microarray, which contains 8,372 unique gene sequences. B, Venn diagrams depicting the number of genes with ≥2-fold changes in mRNA levels. Top, the number of genes induced; bottom, the number of genes repressed by 2.5 and/or 7.5 μmol/L SAHA. C, summary of SAHA-induced alterations in expression of genes involved in androgen signaling. D, real-time PCR analysis of AR, PSA, and kallikrein 2 (KLK2) mRNA levels 2 h following treatment with SAHA.

  • Figure 5.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 5.

    Protein expression changes in LNCaP cells following culture with SAHA. Lysates from cells cultured in the absence or presence of SAHA (2.5, 5, or 7.5 μmol/L) for 12, 24, or 48 h were analyzed by immunoblotting for expression of AR and PSA (A), Her2/neu (B), and cyclin D1 and p21WAF1(C). For each immunoblot, detection of calnexin was used as a loading control (bottom).

  • Figure 6.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 6.

    Effect of SAHA in combination with androgen withdrawal on AR expression, cell proliferation, and cell death. A, LNCaP cells (5 × 105 per well in six-well plates) were cultured for 24 h with SAHA (0, 2.5, or 7.5 μmol/L) in the presence or absence of 5α-dihydrotestosterone (DHT; 1 nmol/L), in either regular RPMI 1640 with 10% FCS (left) or phenol red–free RPMI 1640 with 10% charcoal-stripped FCS (right). Cells were lysed and analyzed by immunoblotting for expression of AR, and calnexin was used as a loading control. B and C, LNCaP cells (2.5 × 104 per well in 24-well plates) were cultured in medium containing charcoal-stripped FCS in the absence or presence of SAHA for up to 7 d. B, cells were counted every 2nd day using a hemocytometer, and cell viability was assessed by trypan blue exclusion. C, the number of dead cells is expressed as a percentage of total cells counted. Points, mean of triplicate wells in a representative experiment; bars, SE.

  • Figure 7.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 7.

    Effect of SAHA in combination with the AR antagonist bicalutamide on LNCaP cell proliferation and cell death. A and B, cells (2.5 × 104 per well in 24-well plates) were cultured in the absence or presence of 0.5 μmol/L SAHA and 1.25 μmol/L bicalutamide, either alone or in combination. A, cells were counted every 2nd day using a hemocytometer, and cell viability was assessed by trypan blue exclusion. B, the number of dead cells is expressed as a percentage of total cells counted. Points, mean of triplicate wells in a representative experiment; bars, SE. C and D, LNCaP cells (2.5 × 104 per well in 24-well plates) were cultured with 0.5 μmol/L SAHA and 1.25 μmol/L bicalutamide, alone or in combination, in the presence or absence of (C) the z-VAD-fmk caspase inhibitor or (D) the androgen 5α-dihydrotestosterone (10 nmol/L) for 5 d. Cell viability was assessed as described above. E, lysates from untreated LNCaP cells (U) and cells cultured with 0.5 μmol/L SAHA (S), 1.25 μmol/L bicalutamide (B), or SAHA and bicalutamide (S+B) were analyzed by immunoblotting for expression of AR and PSA. Calnexin was used as a loading control (bottom).

  • Figure 8.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Figure 8.

    Effect of SAHA in combination with bicalutamide on PC-3 cell proliferation and cell death. PC-3 cells (2.5 × 104 per well in 24-well plates) were cultured in the absence or presence of 0.5 μmol/L SAHA and 1.25 μmol/L bicalutamide, alone or in combination. A, cell viability was assessed as described above. B, the number of dead cells is expressed as a percentage of total cells counted. Points, mean of triplicate wells in a representative experiment; bars, SE.

PreviousNext
Back to top
Molecular Cancer Therapeutics: 6 (1)
January 2007
Volume 6, Issue 1
  • Table of Contents
  • About the Cover

Sign up for alerts

View this article with LENS

Open full page PDF
Article Alerts
Sign In to Email Alerts with your Email Address
Email Article

Thank you for sharing this Molecular Cancer Therapeutics article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
Suberoylanilide hydroxamic acid (vorinostat) represses androgen receptor expression and acts synergistically with an androgen receptor antagonist to inhibit prostate cancer cell proliferation
(Your Name) has forwarded a page to you from Molecular Cancer Therapeutics
(Your Name) thought you would be interested in this article in Molecular Cancer Therapeutics.
Citation Tools
Suberoylanilide hydroxamic acid (vorinostat) represses androgen receptor expression and acts synergistically with an androgen receptor antagonist to inhibit prostate cancer cell proliferation
Deborah L. Marrocco, Wayne D. Tilley, Tina Bianco-Miotto, Andreas Evdokiou, Howard I. Scher, Richard A. Rifkind, Paul A. Marks, Victoria M. Richon and Lisa M. Butler
Mol Cancer Ther January 1 2007 (6) (1) 51-60; DOI: 10.1158/1535-7163.MCT-06-0144

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Share
Suberoylanilide hydroxamic acid (vorinostat) represses androgen receptor expression and acts synergistically with an androgen receptor antagonist to inhibit prostate cancer cell proliferation
Deborah L. Marrocco, Wayne D. Tilley, Tina Bianco-Miotto, Andreas Evdokiou, Howard I. Scher, Richard A. Rifkind, Paul A. Marks, Victoria M. Richon and Lisa M. Butler
Mol Cancer Ther January 1 2007 (6) (1) 51-60; DOI: 10.1158/1535-7163.MCT-06-0144
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • Introduction
    • Materials and Methods
    • Results
    • Discussion
    • Acknowledgments
    • Footnotes
    • References
  • Figures & Data
  • Info & Metrics
  • PDF
Advertisement

Related Articles

Cited By...

More in this TOC Section

  • MAPK-independent impairment of T-cell responses by the multikinase inhibitor sorafenib
  • Interruption of RNA processing machinery by a small compound, 1-[(4-chlorophenyl)methyl]-1H-indole-3-carboxaldehyde (oncrasin-1)
  • Trabectedin (ET-743) promotes differentiation in myxoid liposarcoma tumors
Show more Research Articles: Therapeutics, Targets, and Development
  • Home
  • Alerts
  • Feedback
  • Privacy Policy
Facebook  Twitter  LinkedIn  YouTube  RSS

Articles

  • Online First
  • Current Issue
  • Past Issues
  • Meeting Abstracts

Info for

  • Authors
  • Subscribers
  • Advertisers
  • Librarians
  • Reviewers

About MCT

  • About the Journal
  • Editorial Board
  • Permissions
  • Submit a Manuscript
AACR logo

Copyright © 2019 by the American Association for Cancer Research.

Molecular Cancer Therapeutics
eISSN: 1538-8514
ISSN: 1535-7163

Advertisement