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P2x4 receptor promotes mammary cancer progression by sustaining autophagy and associated mesenchymal transition

Oncogene volume 41, pages 2920–2931 (2022)Cite this article

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Abstract

Metastatic progression is a major burden for breast cancer patients and is associated with the ability of cancer cells to overcome stressful conditions, such as nutrients deprivation and hypoxia, and to gain invasive properties. Autophagy and epithelial-to-mesenchymal transition are critical contributors to these processes. Here, we show that the P2X4 purinergic receptor is upregulated in breast cancer biopsies from patients and it is primarily localised in endolysosomes. We demonstrate that P2X4 enhanced invasion in vitro, as well as mammary tumour growth and metastasis in vivo. The pro-malignant role of P2X4 was mediated by the regulation of lysosome acidity, the promotion of autophagy and cell survival. Furthermore, the autophagic activity was associated with epithelial–to-mesenchymal transition (EMT), and this role of P2X4 was even more pronounced under metabolic challenges. Pharmacological and gene silencing of P2X4 inhibited both autophagy and EMT, whereas its rescue in knocked-down cells led to the restoration of the aggressive phenotype. Together, our results demonstrate a previously unappreciated role for P2X4 in regulating lysosomal functions and fate, promoting breast cancer progression and aggressiveness.

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Fig. 1: Overexpression of P2X4 is associated with mammary tumour growth and metastatic progression.
Fig. 2: P2X4 receptor is partially localised in endolysosomal acidic compartments and controls autophagic flux.
Fig. 3: P2X4 receptor regulates autophagy and promotes cancer cell survival under metabolic challenges.
Fig. 4: P2X4 receptor regulates lysosomal exocytosis and invasive capacities.
Fig. 5: P2X4 drives mammary cancer cells towards a mesenchymal phenotype, associated with increased autophagy.

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All data and materials used in this study are available upon request.

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Acknowledgements

This work was supported by the “Ministère de la Recherche et des Technologies”, the “Ligue Nationale Contre le Cancer – Interrégion Grand-Ouest” to SR, the Région Centre-Val de Loire (grant “CanalEx” to SR), the Institut National du Cancer (grant INCA_16110 “PURIN4EXO” to SR). SR was recipient of a prize “Prix Ruban Rose Avenir 2017” from the Charity “le Cancer du sein: parlons-en!”. SC was recipient of a post-doctoral grant from the School of Life Science, University of Sussex (UK) and from the AURCIL Association in Tours (France). We thank Mrs Carole Desplanches for secretary and administrative assistance. We thank Mrs Stéphanie Rétif, Mrs Marilyne Le Mée and M. Julien Sobilo for the assistance with in vivo experiments performed at CNRS UPS44 CIPA, Orléans. Our microscopy data were obtained with the assistance of the IBiSA Electron Microscopy Facility of the University of Tours, and we are grateful to M. Julien Burlaud-Gaillard for his help with electronic and confocal microscopy. We thank Dr Vinh Ta Phuoc for the insightful discussions on lysosomal distribution analyses. This work was supported by the “Ministère de la Recherche et des Technologies”, the “Ligue Nationale Contre le Cancer – Interrégion Grand-Ouest” to SR, the Région Centre-Val de Loire (grant “CanalEx” to SR), the Institut National du Cancer (grant INCA_16110 “PURIN4EXO” to SR). SR was recipient of a prize “Prix Ruban Rose Avenir 2017” from the Charity “le Cancer du sein: parlons-en!”. SC was recipient of a post-doctoral grant from the School of Life Science, University of Sussex (UK) and from the AURCIL Association in Tours (France).

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Author notes

  1. These authors contributed equally: Ruth Murrell-Lagnado, Sébastien Roger.

Authors and Affiliations

  1. University of Tours, EA4245 Transplantation, Immunology, Inflammation, Tours, France

    Stéphanie Chadet, Jordan Allard, Osbaldo Lopez-Charcas, Roxane Lemoine, Audrey Heraud, Denis Angoulvant & Sébastien Roger

  2. Inserm UMR1069, Nutrition, Growth and Cancer, University of Tours, Tours, France

    Lucie Brisson, Roseline Guibon & Gaëlle Fromont

  3. CNRS UPS44 TAAM, Centre d’Imagerie du Petit animal, Orléans, France

    Stéphanie Lerondel & Alain Le Pape

  4. School of Biomedical Sciences, University of Leeds, Leeds, LS2 9JT, UK

    Lin-Hua Jiang

  5. Sino-UK Joint Laboratory of Brain Function and Injury and Department of Physiology and Pathophysiology, Xinxiang Medical University, Xinxiang, 453003, China

    Lin-Hua Jiang

  6. School of Life Sciences, University of Sussex, Brighton, UK

    Ruth Murrell-Lagnado

  7. Institut Universitaire de France, Paris, France

    Sébastien Roger

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  1. Stéphanie Chadet
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Contributions

All authors contributed extensively to the work presented in this study. SC performed cell culture, molecular and cellular biology experiments, assessed cell viability, apoptosis and invasion, immunofluorescence/confocal imaging, flow cytometry, pH imaging, bioinformatics, and statistical analyses. JA performed invasion and molecular biology experiments. LB and RL participated in cell culture, flow cytometry and scientific input regarding autophagy analysis. AH and JA participated in cell culture. OL-C, JA, and L-HJ participated to rescue experiments. RG and GF performed IHC analyses. SC, SL, and ALP performed in vivo mouse experiments. SC, RM-L, and SR analysed in vivo data. SR and RM-L obtained research grants. DA and L-HJ participated in critical reading of the paper. SC, SR, and RM-L directed the research, designed the study, analysed the data, and wrote the paper.

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Correspondence to Stéphanie Chadet.

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Chadet, S., Allard, J., Brisson, L. et al. P2x4 receptor promotes mammary cancer progression by sustaining autophagy and associated mesenchymal transition. Oncogene 41, 2920–2931 (2022). https://doi.org/10.1038/s41388-022-02297-8

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