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Macroautophagy inhibition sensitizes tamoxifen-resistant breast cancer cells and enhances mitochondrial depolarization

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Abstract

Macroautophagy (autophagy), a process for lysosomal degradation of organelles and long-lived proteins, has been linked to various pathologies including cancer and to the cellular response to anticancer therapies. In the human estrogen receptor positive MCF7 breast adenocarcinoma cell line, treatment with the endocrine therapeutic tamoxifen was shown previously to induce cell cycle arrest, cell death, and autophagy. To investigate specifically the role of autophagy in tamoxifen treated breast cancer cell lines, we used a siRNA approach, targeting three different autophagy genes, Atg5, Beclin-1, and Atg7. We found that knockdown of autophagy, in combination with tamoxifen in MCF7 cells, results in decreased cell viability concomitant with increased mitochondrial-mediated apoptosis. The combination of autophagy knockdown and tamoxifen treatment similarly resulted in reduced cell viability in the breast cancer cell lines, estrogen receptor positive T-47D and tamoxifen-resistant MCF7-HER2. Together, these results indicate that autophagy has a primary pro-survival role following tamoxifen treatment, and suggest that autophagy knockdown may be useful in a combination therapy setting to sensitize breast cancer cells, including tamoxifen-resistant breast cancer cells, to tamoxifen therapy.

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Abbreviations

Atg:

Autophagy-related

BECN1:

Beclin-1

ER:

Estrogen receptor

ER+:

Estrogen receptor positive

GFP:

Green fluorescent protein

HCS:

High content screening

LTGY:

Lysotracker green

HER-2:

Human epidermal growth factor receptor-2

MAP1LC3B or LC3:

Microtubule-associated protein 1 light chain 3 beta

MDC:

Monodansylcadaverine

MMP:

Mitochondrial membrane potential

PR:

Progesterone receptor

RNAi:

RNA interference

SERM:

Selective estrogen receptor modulator

SiRNA:

Small interfering RNA

TAM:

Tamoxifen

TMZ:

Temozolomide

3-MA:

3-Methyladenine

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Acknowledgements

We thank T. Yoshimori for the GFP-LC3 plasmid, M. Roberge, I. Nabi, A. Balgi and P. Lajoie for the stably transfected MCF7-GFP-LC3 cell line, J. Halaschek-Wiener and M. Marra for helpful comments on the manuscript, Steven Poon for valuable suggestions in designing the segmentation protocols for high content screening, S. Oomah for assistance with primer design, and the British Columbia Cancer Foundation for support. SMG also thanks S. O’Reilly for assistance through an Abbott Research Grant. This work was supported by USAMRMC Breast Cancer Research Program Concept Award Number W81XWH-05-1-04 and Canadian Breast Cancer Foundation BC/Yukon Chapter grant to SMG. The Canadian Breast Cancer Research Alliance and Canadian Institutes of Health Research also supported aspects of these studies (MBB and WHD).

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Authors and Affiliations

  1. Genome Sciences Centre, British Columbia Cancer Agency, 675 West 10th Avenue, Vancouver, BC, Canada, V5Z 1L3

    M. A. Qadir, B. Kwok, K. H. To, D. Le & Sharon M. Gorski

  2. Department of Advanced Therapeutics, British Columbia Cancer Agency, 600 West 10th Avenue, Vancouver, BC, Canada, V5Z 4E6

    W. H. Dragowska & M. B. Bally

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  1. M. A. Qadir
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  2. B. Kwok
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Correspondence to Sharon M. Gorski.

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Qadir, M.A., Kwok, B., Dragowska, W.H. et al. Macroautophagy inhibition sensitizes tamoxifen-resistant breast cancer cells and enhances mitochondrial depolarization. Breast Cancer Res Treat 112, 389–403 (2008). https://doi.org/10.1007/s10549-007-9873-4

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