Induction of cell cycle arrest and DNA damage by the HDAC inhibitor panobinostat (LBH589) and the lipid peroxidation end product 4-hydroxynonenal in prostate cancer cells

doi:10.1016/j.freeradbiomed.2010.11.011

Free Radical Biology and Medicine

Volume 50, Issue 2, 15 January 2011, Pages 313-322

https://doi.org/10.1016/j.freeradbiomed.2010.11.011 Get rights and content

Abstract

Histone deacetylase inhibitors (HDACIs) are promising antineoplastic agents for the treatment of cancer. Here we report that the lipid peroxidation end product 4-hydroxynonenal (HNE) significantly potentiates the anti-tumor effects of the HDAC inhibitor panobinostat (LBH589) in the PC3 prostate cancer cell model. Panobinostat and HNE inhibited proliferation of PC3 cells and the combination of the two agents resulted in a significant combined effect. Cell cycle analysis revealed that both single agents and, to a greater extent, their combined treatment induced G2/M arrest, but cell death occurred in the combined treatment only. Furthermore, HNE and, to a greater extent, the combined treatment induced dephosphorylation of Cdc2 leading to progression into mitosis as confirmed by α-tubulin/DAPI staining and phospho-histone H3 (Ser10) analysis. To evaluate possible induction of DNA damage we utilized the marker phosphorylated histone H2A.X. Results showed that the combination of panobinostat and HNE induced significant DNA damage concomitant with the mitotic arrest. Then, by using androgen receptor (AR)-expressing PC3 cells we observed that the responsiveness to HNE and panobinostat was independent of the expression of functional AR. Taken together, our data suggest that HNE potentiates the antitumoral effect of the HDACI panobinostat in prostate cancer cells.

Section snippets

Compounds

Panobinostat (LBH589) was generously provided by Novartis Pharmaceutical and was dissolved in dimethyl sulfoxide (DMSO). HNE was purchased from Cayman Chemicals (32100) and was prepared as follows: HNE was dissolved in ethanol that was evaporated through a gentle flow of N₂ and subsequently resuspended in sterile phosphate-buffered saline (PBS). The concentration was measured by spectrophotometer recording the absorbance of an aliquot of HNE diluted 1:200 in water at 223 nm (ε = 13,750).

Primary antibodies

The

Results

The effects of panobinostat on prostate cell proliferation and viability are reported in Fig. 1. Panobinostat reduced the growth of all three prostate cancer cell lines LNCaP, DU145, and PC3, in a dose-dependent manner. However, the panobinostat-mediated inhibition of proliferation was significantly more pronounced in LNCaP and DU145 compared to PC3 cells. Moreover, panobinostat-mediated antiproliferative effect was associated with significant dose-dependent reduction of cell viability,

Discussion

Our results suggest that nanomolar concentrations of the HDAC inhibitor panobinostat have significant anti-tumor activity in several prostate cancer cell lines. However, the sensitivity varies and may involve either induction of cell death, as observed in LNCaP and DU145 cells, or growth inhibition as demonstrated in PC3 cells. In the latter, the antiproliferative efficacy of panobinostat was also significantly less pronounced in comparison to LNCaP and DU145 cells. Similar results have been

Acknowledgments

This work was supported by Compagnia di San Paolo (MIUR-PRIN 2004 2004062075 to G.B.) and the National Cancer Institute (P50 CA58236 to R.P.).

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¹: These authors contributed equally to this work.

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Original ContributionInduction of cell cycle arrest and DNA damage by the HDAC inhibitor panobinostat (LBH589) and the lipid peroxidation end product 4-hydroxynonenal in prostate cancer cells

Abstract

Section snippets

Compounds

Primary antibodies

Results

Discussion

Acknowledgments

Blood

Cancer Lett.

Free Radic. Biol. Med.

Free Radic. Biol. Med.

Mol. Aspects Med.

Free Radic. Biol. Med.

J. Nutr. Biochem.

Exp. Cell Res.

Free Radic. Biol. Med.

J. Lipid Mediatiators Cell Signal.

Free Radic. Biol. Med.

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Free Radic. Biol. Med.

Neoplasia

American Cancer Society cancer statistics

CA Cancer J. Clin.

The role of histone deacetylases in prostate cancer

Epigenetics

Histone deacetylase inhibitors in cancer therapy

J. Clin. Oncol.

Histone deacetylase inhibitors and the promise of epigenetic (and more) treatments for cancer

Nat. Rev. Cancer

Role of histone deacetylase inhibitor-induced reactive oxygen species and DNA damage in LAQ-824/fludarabine antileukemic interactions

Mol. Cancer Ther.

Original Contribution
Induction of cell cycle arrest and DNA damage by the HDAC inhibitor panobinostat (LBH589) and the lipid peroxidation end product 4-hydroxynonenal in prostate cancer cells