Carnosol, a Dietary Diterpene, Displays Growth Inhibitory Effects in Human Prostate Cancer PC3 Cells Leading to G2-Phase Cell Cycle Arrest and Targets the 5′-AMP-Activated Protein Kinase (AMPK) Pathway (original) (raw)

Abstract

Purpose

This study examines the anti-cancer effect of carnosol in human prostate cancer PC3 cells and its role in modulating multiple signaling pathways associated with carcinogenesis.

Methods

PC3 cells were treated with carnosol and were evaluated using a flow cytometry, a protein array and Western blot analysis to identify signaling pathways targeted by carnosol.

Results

Using an MTT assay we found that carnosol (10–70 μM) decreases cell viability in a time and dose-dependent manner. Further analysis using flow cytometry as well as biochemical analysis identified G2-phase cell cycle arrest. To establish a more precise mechanism, we performed a protein array that evaluated 638 proteins involved in cell signaling pathways. The protein array identified 5′-AMP-activated protein kinase (AMPK), a serine/threonine protein kinase involved in the regulation of cellular energy balance as a potential target. Further downstream effects consistent with cancer inhibition included the modulation of the mTOR/HSP70S6k/4E-BP1 pathway. Additionally, we found that carnosol targeted the PI3K/Akt pathway in a dose dependent manner.

Conclusions

These results suggest that carnosol targets multiple signaling pathways that include the AMPK pathway. The ability of carnosol to inhibit prostate cancer in vitro suggests carnosol may be a novel agent for the management of PCa.

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Abbreviations

AICAR:

5-aminoimidazole-4-carboxamide ribonucleoside

AMPK:

5′-AMP-activated protein kinase

cdks:

cyclin dependent kinases

DMSO:

dimethyl sulfoxide

FBS:

fetal bovine serum

LH-RH:

luteinizing hormone-releasing hormone

mTOR:

mammalian target of rapamycin

PBS:

phophsphate buffered saline

Pca:

prostate cancer

PI3K:

phosphatidylinositol 3-kinase

PTEN:

phosphatase and tensin homologue deleted on chromosome ten

TSC2:

tuberous sclerosis complex 2

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Acknowledgements

This work was supported by a Clinical and Translational Science Award (CTSA) training grant (J.Johnson) through the Institute for Clinical and Translation Research (ICTR) at the University of Wisconsin. (NIH 1KL2RR025012-01 and K12 RR023268).

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Authors and Affiliations

  1. Center for Women’s Health Research, University of Wisconsin, School of Medicine and Public Health, Madison, Wisconsin, USA
    Jeremy J. Johnson & Chenelle R. Heren
  2. Department of Dermatology, University of Wisconsin, School of Medicine and Public Health, Madison, Wisconsin, USA
    Jeremy J. Johnson, Deeba N. Syed, Chenelle R. Heren, Yewseok Suh, Vaqar M. Adhami & Hasan Mukhtar
  3. Department of Dermatology, University of Wisconsin, Medical Sciences Center, Rm. B-25, 1300 University Ave., Madison, Wisconsin, 53706, USA
    Hasan Mukhtar

Authors

  1. Jeremy J. Johnson
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  2. Deeba N. Syed
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  3. Chenelle R. Heren
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  4. Yewseok Suh
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  5. Vaqar M. Adhami
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  6. Hasan Mukhtar
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Corresponding author

Correspondence toHasan Mukhtar.

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Johnson, J.J., Syed, D.N., Heren, C.R. et al. Carnosol, a Dietary Diterpene, Displays Growth Inhibitory Effects in Human Prostate Cancer PC3 Cells Leading to G2-Phase Cell Cycle Arrest and Targets the 5′-AMP-Activated Protein Kinase (AMPK) Pathway.Pharm Res 25, 2125–2134 (2008). https://doi.org/10.1007/s11095-008-9552-0

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