A water-soluble parthenolide analogue suppresses in vivo prostate cancer growth by targeting NFκB and generating reactive oxygen species

Rajasubramaniam Shanmugam, Praveen Kusumanchi, Liang Cheng, Peter Crooks, Sundar Neelakantan, William Matthews, Harikrishna Nakshatri, Christopher J. Sweeney

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

BACKGROUND. To characterize the molecular changes associated with DMAPT-induced prostate cancer cell death and its in vivo activity. METHODS. CWR22Rv1 and PC-3 were subjected to flow cytometry, electrophoretic mobility shift assays, and Western blot studies to measure DMAPT's ability to generate reactive oxygen species (ROS), inhibit NFκB DNA binding, and cause changes in anti-apoptotic proteins. N-acetyl cysteine (NAC) and short hairpin RNA (shRNA) were used to determine the contribution of ROS and JNK2 activation, respectively. The BrdU incorporation assay was used to measure proliferation and trypan blue studies assessed cell viability after DMAPT treatment. The in vivo activity of DMAPT as a single agent and in combination with bicalutamide or docetaxel was assessed in a subcutaneous xenograft model with athymic nude female mice. RESULTS. DMAPT generated ROS with subsequent JNK activation and inhibited NFκB DNA binding and expression of NFκB-regulated anti-apoptotic proteins. DMAPT increased necrotic and apoptotic cell death in a cell-type-dependent manner and both types of cell death were blocked by NAC. Additionally, shRNA JNK2 partially blocked the anti-proliferative activity of DMAPT. DMAPT inhibited CWR22Rv1 and PC-3 cellular proliferation by 100% with 10 and 20 μM respectively and in vivo, DMAPT was more effective at inhibiting growth than biclutamide (CWR22v1) and docetaxel (PC-3). CONCLUSIONS. DMAPT promotes cell death by both generating ROS and inhibition of NFκB. Its in vivo activity supports the conduct of clinical trials in patients with castrate-resistant disease.

Original languageEnglish
Pages (from-to)1074-1086
Number of pages13
JournalProstate
Volume70
Issue number10
DOIs
StatePublished - Jul 1 2010

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docetaxel
Reactive Oxygen Species
Prostatic Neoplasms
Cell Death
Apoptosis Regulatory Proteins
Water
Growth
Nude Mice
Small Interfering RNA
Cysteine
Trypan Blue
DNA
Electrophoretic Mobility Shift Assay
Bromodeoxyuridine
Heterografts
Cell Survival
Flow Cytometry
Western Blotting
Cell Proliferation
Clinical Trials

Keywords

  • Androgen independence
  • Apoptosis
  • Parthenolide analogue

ASJC Scopus subject areas

  • Urology
  • Oncology
  • Medicine(all)

Cite this

A water-soluble parthenolide analogue suppresses in vivo prostate cancer growth by targeting NFκB and generating reactive oxygen species. / Shanmugam, Rajasubramaniam; Kusumanchi, Praveen; Cheng, Liang; Crooks, Peter; Neelakantan, Sundar; Matthews, William; Nakshatri, Harikrishna; Sweeney, Christopher J.

In: Prostate, Vol. 70, No. 10, 01.07.2010, p. 1074-1086.

Research output: Contribution to journalArticle

Shanmugam, Rajasubramaniam ; Kusumanchi, Praveen ; Cheng, Liang ; Crooks, Peter ; Neelakantan, Sundar ; Matthews, William ; Nakshatri, Harikrishna ; Sweeney, Christopher J. / A water-soluble parthenolide analogue suppresses in vivo prostate cancer growth by targeting NFκB and generating reactive oxygen species. In: Prostate. 2010 ; Vol. 70, No. 10. pp. 1074-1086.
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T1 - A water-soluble parthenolide analogue suppresses in vivo prostate cancer growth by targeting NFκB and generating reactive oxygen species

AU - Shanmugam, Rajasubramaniam

AU - Kusumanchi, Praveen

AU - Cheng, Liang

AU - Crooks, Peter

AU - Neelakantan, Sundar

AU - Matthews, William

AU - Nakshatri, Harikrishna

AU - Sweeney, Christopher J.

PY - 2010/7/1

Y1 - 2010/7/1

N2 - BACKGROUND. To characterize the molecular changes associated with DMAPT-induced prostate cancer cell death and its in vivo activity. METHODS. CWR22Rv1 and PC-3 were subjected to flow cytometry, electrophoretic mobility shift assays, and Western blot studies to measure DMAPT's ability to generate reactive oxygen species (ROS), inhibit NFκB DNA binding, and cause changes in anti-apoptotic proteins. N-acetyl cysteine (NAC) and short hairpin RNA (shRNA) were used to determine the contribution of ROS and JNK2 activation, respectively. The BrdU incorporation assay was used to measure proliferation and trypan blue studies assessed cell viability after DMAPT treatment. The in vivo activity of DMAPT as a single agent and in combination with bicalutamide or docetaxel was assessed in a subcutaneous xenograft model with athymic nude female mice. RESULTS. DMAPT generated ROS with subsequent JNK activation and inhibited NFκB DNA binding and expression of NFκB-regulated anti-apoptotic proteins. DMAPT increased necrotic and apoptotic cell death in a cell-type-dependent manner and both types of cell death were blocked by NAC. Additionally, shRNA JNK2 partially blocked the anti-proliferative activity of DMAPT. DMAPT inhibited CWR22Rv1 and PC-3 cellular proliferation by 100% with 10 and 20 μM respectively and in vivo, DMAPT was more effective at inhibiting growth than biclutamide (CWR22v1) and docetaxel (PC-3). CONCLUSIONS. DMAPT promotes cell death by both generating ROS and inhibition of NFκB. Its in vivo activity supports the conduct of clinical trials in patients with castrate-resistant disease.

AB - BACKGROUND. To characterize the molecular changes associated with DMAPT-induced prostate cancer cell death and its in vivo activity. METHODS. CWR22Rv1 and PC-3 were subjected to flow cytometry, electrophoretic mobility shift assays, and Western blot studies to measure DMAPT's ability to generate reactive oxygen species (ROS), inhibit NFκB DNA binding, and cause changes in anti-apoptotic proteins. N-acetyl cysteine (NAC) and short hairpin RNA (shRNA) were used to determine the contribution of ROS and JNK2 activation, respectively. The BrdU incorporation assay was used to measure proliferation and trypan blue studies assessed cell viability after DMAPT treatment. The in vivo activity of DMAPT as a single agent and in combination with bicalutamide or docetaxel was assessed in a subcutaneous xenograft model with athymic nude female mice. RESULTS. DMAPT generated ROS with subsequent JNK activation and inhibited NFκB DNA binding and expression of NFκB-regulated anti-apoptotic proteins. DMAPT increased necrotic and apoptotic cell death in a cell-type-dependent manner and both types of cell death were blocked by NAC. Additionally, shRNA JNK2 partially blocked the anti-proliferative activity of DMAPT. DMAPT inhibited CWR22Rv1 and PC-3 cellular proliferation by 100% with 10 and 20 μM respectively and in vivo, DMAPT was more effective at inhibiting growth than biclutamide (CWR22v1) and docetaxel (PC-3). CONCLUSIONS. DMAPT promotes cell death by both generating ROS and inhibition of NFκB. Its in vivo activity supports the conduct of clinical trials in patients with castrate-resistant disease.

KW - Androgen independence

KW - Apoptosis

KW - Parthenolide analogue

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