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Regulation of apoptosis by long non-coding RNA GAS5 in breast cancer cells: implications for chemotherapy

  • Preclinical Study
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Abstract

The putative tumour suppressor and apoptosis-promoting gene, growth arrest-specific 5 (GAS5), encodes long ncRNA (lncRNA) and snoRNAs. Its expression is down-regulated in breast cancer, which adversely impacts patient prognosis. In this preclinical study, the consequences of decreased GAS5 expression for breast cancer cell survival following treatment with chemotherapeutic agents are addressed. In addition, functional responses of triple-negative breast cancer cells to GAS5 lncRNA are examined, and mTOR inhibition as a strategy to enhance cellular GAS5 levels is investigated. Breast cancer cell lines were transfected with either siRNA to GAS5 or with a plasmid encoding GAS5 lncRNA and the effects on breast cancer cell survival were determined. Cellular responses to mTOR inhibitors were evaluated by assaying culture growth and GAS5 transcript levels. GAS5 silencing attenuated cell responses to apoptotic stimuli, including classical chemotherapeutic agents; the extent of cell death was directly proportional to cellular GAS5 levels. Imatinib action in contrast, was independent of GAS5. GAS5 lncRNA promoted the apoptosis of triple-negative and oestrogen receptor-positive cells but only dual PI3K/mTOR inhibition was able to enhance GAS5 levels in all cell types. Reduced GAS5 expression attenuates apoptosis induction by classical chemotherapeutic agents in breast cancer cells, providing an explanation for the relationship between GAS5 expression and breast cancer patient prognosis. Clinically, this relationship may be circumvented by the use of GAS5-independent drugs such as imatinib, or by restoration of GAS5 expression. The latter may be achieved by the use of a dual PI3K/mTOR inhibitor, to improve apoptotic responses to conventional chemotherapies.

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Abbreviations

A490:

Absorbance at 490 nm

C T :

Threshold cycle

EST:

Expressed sequence tag

5-FU:

5-Fluorouracil

GAS5:

Growth arrest-specific 5

HER2:

Human epidermal growth factor receptor 2

lncRNA:

Long ncRNA

MCT:

Multiple comparison test

NC:

Negative control

ncRNA:

Non-coding RNA

NMD:

Nonsense-mediated decay

RT-qPCR:

Real time reverse transcription polymerase chain reaction

5′-TOP:

5′-Terminal oligopyrimidine tract

TNBC:

Triple-negative breast cancer

UV-C:

Ultraviolet-C

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Acknowledgment

This study was supported by a grant from the Breast Cancer Campaign, which is gratefully acknowledged.

Conflict of interest

The authors declare that they have no conflicts of interest.

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Authors and Affiliations

  1. Apoptosis Research Group, Institute of Science and Technology in Medicine, School of Life Sciences, Keele University, Huxley Building, Keele, ST5 5BG, UK

    Mark R. Pickard & Gwyn T. Williams

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  1. Mark R. Pickard
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  2. Gwyn T. Williams
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Correspondence to Mark R. Pickard or Gwyn T. Williams.

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Pickard, M.R., Williams, G.T. Regulation of apoptosis by long non-coding RNA GAS5 in breast cancer cells: implications for chemotherapy. Breast Cancer Res Treat 145, 359–370 (2014). https://doi.org/10.1007/s10549-014-2974-y

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  • DOI: https://doi.org/10.1007/s10549-014-2974-y

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