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CCL8 mediates crosstalk between endothelial colony forming cells and triple-negative breast cancer cells through IL-8, aggravating invasion and tumorigenicity

Abstract

Triple-negative breast cancer (TNBC) is an aggressive type of breast cancer with a poor prognosis for which no effective therapeutic measures are currently available. The present study aimed to investigate whether interactions with endothelial colony-forming cells (ECFCs) promote aggressive progression of TNBC cells. Herein, using an indirect co-culture system, we showed that co-culture increased the invasive and migratory phenotypes of both MDA-MB-231 TNBC cells and ECFCs. Through a cytokine antibody array and RT-PCR analysis, we revealed that co-culture markedly induced secretion of the chemokine C-C motif ligand (CCL)8 from ECFCs and that of interleukin (IL)-8 from MDA-MB-231 cells. CCL8 was crucial for ECFC-induced IL-8 secretion and invasion of MDA-MB-231 cells as well as for MDA-MB-231-enhanced MMP-2 secretion and angiogenesis of ECFCs. We suggest c-Jun as a transcription factor for CCL8-induced IL-8 expression in MDA-MB-231 cells. IL-8 was important for co-culture-induced CCL8 and MMP-2 upregulation and invasion of ECFCs. Notably, our findings reveal a positive feedback loop between CCL8 and IL-8, which contributes to the aggressive phenotypes of both ECFC and TNBC cells. Using an MDA-MB-231 cell-based xenograft model, we show that tumor growth and metastasis are increased by co-injected ECFCs in vivo. Increased expression of IL-8 was observed in tissues with bone metastases in mice injected with conditioned media from co-cultured cells. High IL-8 levels are correlated with poor recurrence-free survival in TNBC patients. Together, these results suggest that CCL8 and IL-8 mediate the crosstalk between ECFCs and TNBC, leading to aggravation of tumorigenicity in TNBC.

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Fig. 1: ECFCs increase proliferation, invasion, migration, and anchorage-independent growth in MDA-MB-231 TNBC cells.
Fig. 2: The invasive, migratory, and angiogenic properties and MMP-2 expression of ECFCs are enhanced by co-culture with MDA-MB-231 cells.
Fig. 3: Co-culture induces secretion of IL-8 from MDA-MB-231 cells and secretion of CCL8 and PDGF-BB from ECFCs.
Fig. 4: CCL8 is crucial for ECFC-induced IL-8 expression and invasion of MDA-MB-231 cells as well as MDA-MB-231-enhanced MMP-2 and angiogenesis of ECFCs.
Fig. 5: c-Jun is involved in transcription of IL-8 induced by ECFCs and CCL8.
Fig. 6: Tumor growth, metastasis, and expression of CCL8 and IL-8 are increased by ECFCs in vivo.

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Acknowledgements

The present study was supported by the National Research Foundation of Korea (no. 2016R1A6A1A03007648 and no. 2019R1A2C1009773).

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Kim, ES., Nam, SM., Song, H.K. et al. CCL8 mediates crosstalk between endothelial colony forming cells and triple-negative breast cancer cells through IL-8, aggravating invasion and tumorigenicity. Oncogene 40, 3245–3259 (2021). https://doi.org/10.1038/s41388-021-01758-w

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