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Hypoxia Induces the Acquisition of Cancer Stem-like Phenotype Via Upregulation and Activation of Signal Transducer and Activator of Transcription-3 (STAT3) in MDA-MB-231, a Triple Negative Breast Cancer Cell Line

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Cancer Microenvironment

Abstract

The finding that hypoxia can induce cancer stemness in various experimental models is in agreement with the conceptual basis of cancer cell plasticity. Here, we aimed to gain insights into the molecular basis of hypoxia-induced cancer cell plasticity in triple negative breast cancer (TNBC). To achieve this goal, we employed our previously published in-vitro model of TNBC, in which a small subset of stem-like cells can be distinguished from the bulk cell population based on their responsiveness to a Sox2 reporter. In MDA-MB-231, a TNBC cell line, we observed that hypoxia significantly increased the expression of luciferase and green fluorescence protein (GFP), the readouts of the Sox2 reporter. Upon hypoxic challenge, the bulk, reporter unresponsive (RU) cells acquired stem-like features, as evidenced by the significant increases in the proportion of CD44high/CD24low cells, colony formation and resistance to cisplatin. Correlating with these phenotypic changes, RU cells exposed to hypoxia exhibited a substantial upregulation of the active/phosphorylated form of STAT3 (pSTAT3). This hypoxia-induced activation of STAT3 correlated with increased STAT3 transcriptional activity, as evidenced by increased STAT3-DNA binding and an altered gene expression profile. This hypoxia-induced STAT3 activation is biologically significant, since siRNA knockdown of STAT3 in RU cells significantly attenuated the hypoxia-induced acquisition of Sox2 activity and stem-like phenotypic features. In conclusion, our data have provided the proof-of-concept that STAT3 is a critical mediator in promoting the hypoxia-induced acquisition of cancer stemness in TNBC. Targeting STAT3 in TNBC may be useful in overcoming chemoresistance and decreasing the risk of disease relapse.

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Acknowledgements

This work was financially supported by grants from Canadian Institutes of Health Research (CIHR) MOP 137153 and Canadian Breast Cancer Foundation (CBCF) awarded to A.L and R.L. H.S.A was awarded the Women and Children’s Health Research Institute (WCHRI) and Alberta Cancer Foundation (ACF) Graduate Studentships. N.G was funded by CBCF. The authors would like to thank Amir Soleimani, Department of Pharmacy and Pharmaceutical Sciences, University of Alberta, for critical reading of the manuscript.

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Authors and Affiliations

  1. Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, AB, T6G 2E1, Canada

    Hoda Soleymani Abyaneh & Afsaneh Lavasanifar

  2. Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, AB, T6G 2E1, Canada

    Nidhi Gupta, Abdulraheem Alshareef, Keshav Gopal & Raymond Lai

  3. Department of Chemical & Materials Engineering, Faculty of Engineering, University of Alberta, Edmonton, AB, T6G 1H9, Canada

    Afsaneh Lavasanifar

  4. Department of Oncology, University of Alberta, Edmonton, AB, T6G 2E1, Canada

    Raymond Lai

  5. DynaLIFEDx Medical Laboratories, Edmonton, AB, T5J 5E2, Canada

    Raymond Lai

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  1. Hoda Soleymani Abyaneh
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  2. Nidhi Gupta
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  3. Abdulraheem Alshareef
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  6. Raymond Lai
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Contributions

H.S.A designed the research plan, carried out experiments and wrote the manuscript. N.G contributed to the design and performance of the experiments and data analysis and critical reading of the manuscript. A.A contributed to the design and data analysis of oligonucleotide arrays experiment and critical reading of the manuscript. K.G assisted with the flow cytometric detection of RU/RR conversion. A.L and R.L conceived and designed the research plan and critical reading of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Raymond Lai.

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Soleymani Abyaneh, H., Gupta, N., Alshareef, A. et al. Hypoxia Induces the Acquisition of Cancer Stem-like Phenotype Via Upregulation and Activation of Signal Transducer and Activator of Transcription-3 (STAT3) in MDA-MB-231, a Triple Negative Breast Cancer Cell Line. Cancer Microenvironment 11, 141–152 (2018). https://doi.org/10.1007/s12307-018-0218-0

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