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A new synthetic HDAC inhibitor, MHY218, induces apoptosis or autophagy-related cell death in tamoxifen-resistant MCF-7 breast cancer cells

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Summary

Acquired resistance to tamoxifen (Tam) is a critical problem in breast cancer therapy. Therefore, new potential strategies for Tam-resistant breast cancer are needed recently. In this study, we synthesized a novel histone deacetylase (HDAC) inhibitor, MHY218, for the development of potent inhibitors of HDAC and evaluated its biological activities by monitoring the anticancer effects in Tam-resistant MCF-7 (TAMR/MCF-7) cells via in vitro and in vivo studies. MHY218 significantly inhibited the proliferation of TAMR/MCF-7 cells in a dose-dependent manner. The total HDAC enzyme activity was significantly inhibited, corresponding with inhibition of acetylated H3 and H4 expression in TAMR/MCF-7 cells. HDAC1, 4, and 6 expression levels were decreased in response to MHY218 treatment. Cell cycle analysis indicated that MHY218 induced G2/M phase cell cycle arrest. As expected, apoptotic cell death was observed in response to MHY218 treatment. Interestingly, levels of beclin-1 and LC3-II, the markers of autophagy, were increased in TAMR/MCF-7 cells treated with MHY218. The efficacy of MHY218 was also compared with that of SAHA in vivo in a xenograft model of nude mice bearing a TAMR/MCF-7 cells. MHY218 (10 mg/kg, twice a week for 21 days) completely inhibited tumor growth and MHY218 markedly inhibited the expression of proliferative cell nuclear antigen (PCNA) in tumor tissue. These results indicate that MHY218 can induce caspase-independent autophagic cell death rather than apoptotic cell death. The MHY218-induced autophagic cell death could be a new strategy in the treatment of Tam-resistant human breast cancer.

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Abbreviations

Tam:

Tamoxifen

HDAC:

Histone deacetylase

SAHA:

Suberoylanilide hydroxamic acid

DMSO:

Dimethyl sulfoxide

TAMR/MCF-7:

Tamoxifen-resistant MCF-7

EBCTCG:

Early Breast Cancer Trialists’ Collaborative Groups

PKA:

Protein kinase A

FBS:

Fetal bovine serum

SDS:

Sodium dodecylsulfate

MTT:

3(4,5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide N-acetylcysteine

PI:

Propidium iodide

PARP:

Poly-ADP ribose polymerase

PBS:

Phosphate-buffered saline

PVDF:

Polyvinylidene difluoride

ECL:

Enhanced chemiluminescence

TsA:

Trichostatin A

DAPI:

4,6-diamidino-2-phenylindole

HRP:

Horseradish peroxidase

IC50 :

50% inhibitory concentration

CDK:

Cyclin-dependent kinase

PCNA:

Proliferative cell nuclear antigen

SDS-PAGE:

Sodium dodecylsulfate gel electrophoresis

SPF:

Specific pathogen free

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Acknowledgements

This work was supported by National Research Foundation of Korea (NRF) grants funded by the Korea Government (No.20090083538 and KRF-2008-314-E00292).

Conflict of interest

No financial and personal conflicts of interest were disclosed.

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Authors and Affiliations

  1. College of Pharmacy and MRC Center, Pusan National University, San 30, Jangjeon-dong, Geumjeong-gu, Busan, 609-735, Republic of Korea

    Ji Hye Park, Mee Young Ahn, Tae Hyung Kim, Jaewon Lee, Hyung Ryong Moon, Jee H. Jung, Hae Young Chung & Hyung Sik Kim

  2. Research Institute, National Cancer Center, 809 Madu 1-dong, Ilsan-gu, Goyang-si, Gyeonggi-do, Republic of Korea

    Sungpill Yoon

  3. College of Pharmacy, Seoul National University, 151-754 Shilim-dong, Kwanak-gu, Seoul, Republic of Korea

    Keon Wook Kang

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  1. Ji Hye Park
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Supplementary Fig. 1

Monodansylcadaverine (MDC) staining shows autophagy was activated in TAMR/MCF-7 cells after treatment with MHY218. Cultured cells were incubated with indicated concentration of drugs for 24 h, fixed with 3.7% paraformaldehyde and then stained with 0.05 mM MDC. Cells were examined by Zeiss confocal microscope LSM 510. Scale bar = 10 μm. Fluorescence particles with blue dots show autophagic vacuoles. The data shown are representative of three independent experiments. (JPEG 174 kb)

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Park, J.H., Ahn, M.Y., Kim, T.H. et al. A new synthetic HDAC inhibitor, MHY218, induces apoptosis or autophagy-related cell death in tamoxifen-resistant MCF-7 breast cancer cells. Invest New Drugs 30, 1887–1898 (2012). https://doi.org/10.1007/s10637-011-9752-z

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