Extracellular ATP promotes breast cancer invasion and chemoresistance via SOX9 signaling

Abstract

Our previous research demonstrated that extracellular adenosine 5′-triphosphate (ATP) could promote breast cancer cell invasion. However, the impact of extracellular ATP on chemoresistance and the mechanisms behind ATP pro-invasion and pro-chemoresistance remain unclear. Here we aimed to determine the molecules or signaling pathways involved. cDNA microarray was performed to identify the differentially expressed genes before and after ATP treatment. As a result, Sex-determining region Y-box 9 (SOX9) was up-regulated after ATP treatment in breast cancer cells. In vitro invasion and migration assays demonstrated that knocking down SOX9 attenuated ATP-driven invasive capability. Mass spectrometry and co-IP revealed that SOX9 interacted with Janus kinase 1 (JAK1). Afterward, IL-6-JAK1-STAT3 signaling was demonstrated to promote SOX9 expression and invasion following ATP treatment. Notably, ATP-IL-6-SOX9 signaling was shown to stimulate chemoresistance in breast cancer cells. ChIP assays identified some potential SOX9 target genes, among which carcinoembryonic antigen-related cell adhesion molecule 5/6 (CEACAM5/6) was demonstrated to mediate ATP pro-invasive function, while ATP-binding cassette subfamily B member 1 (ABCB1) and ATP-binding cassette subfamily G member 2 (ABCG2) mediated ATP-driven chemoresistance. In addition, SOX9-knockdown and apyrase (an ATP hydrolase)-treated MDA-MB-231 cells illustrated decreased tumor growth and enhanced drug sensitivity in nude mice. In vitro spheroid formation assays also proved the significance of ATP-SOX9 in mediating chemoresistance. Moreover, molecules involved in ATP-SOX9 signaling were up-regulated in human breast carcinoma specimens and were associated with poor prognosis. Altogether, SOX9 signaling is vital in ATP-driven invasion and chemoresistance, which may serve as a potential target for breast cancer therapies.

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Fig. 1: The up-regulation of SOX9 contributes to ATP-promoted breast cancer cell invasion in vitro.
Fig. 2: IL-6-JAK1-STAT3 is involved in ATP-SOX9 signaling and cancer cell invasion.
Fig. 3: IL-6-JAK1-STAT3-SOX9 axis is required for ATP-driven chemoresistance in breast cancer cells.
Fig. 4: ATP promotes invasion and chemoresistance via SOX9 targets CEACAM5/6, ABCB1, and ABCG2, respectively.
Fig. 5: ATP-SOX9 signaling promotes xenograft tumor growth, metastasis, and chemoresistance in vivo.
Fig. 6: ATP-SOX9 signaling mediates spheroid formation and chemoresistance in vitro.
Fig. 7: ATP-SOX9 axis is associated with clinical breast cancer progression.
Fig. 8: The proposed model for the ATP-SOX9 signaling.

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Acknowledgements

This work was supported by grants to XXT and WGF from the National Natural Science Foundation of China (No 81872382 and 81621063).

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Correspondence to Wei-Gang Fang or Xin-Xia Tian.

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Yang, H., Geng, Y., Wang, P. et al. Extracellular ATP promotes breast cancer invasion and chemoresistance via SOX9 signaling. Oncogene (2020). https://doi.org/10.1038/s41388-020-01402-z

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