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Interleukin 17A promotes cell migration, enhances anoikis resistance, and creates a microenvironment suitable for triple negative breast cancer tumor metastasis

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Abstract

Background

The aim of this study was to investigate the role of IL-17A in the cancer microenvironment and the recurrence of triple negative breast cancer (TNBC).

Methods

Using human TNBC cell lines, the role of IL17-A was investigated by knocked down of IL-17A (ΔIL-17A) and by administration of IL-17A into the culture medium. Cell proliferation assays, migration assays, as well as Western blot analysis and real-time PCR, were used to evaluate IL-17A-related signaling. Three types of 4T1 cells were implanted into BALB/c mice, namely wild type (WT), ΔIL-17A, and WT + neutralizing IL-17 antibody (WT + Ab) cells. Tumor weight, necrosis area, and the number of circulating tumor cells (CTCs) were measured. Immunohistochemistry and Western blotting were used to analyze expression of CD34, CD8, and TGF-β1 as well as anoikis resistance. The Kaplan–Meier’s method was used to correlate IL-17A expression and patient outcome, including disease-free survival (DFS) and overall survival (OS).

Results

Our results demonstrated that IL-17A was able to stimulate the migratory activity, but not the growth rate, of MDA-MB-231/468 cells. In vivo, for the ΔIL-17A group, there was an increase in necrosis area, a decrease in tumor CD34 expression and a reduction in the number of CTCs. Furthermore, in WT + Ab group, there was a decreased in tumor expression of CD34, fewer CD8 ( +) cells, and fewer CTCs, but an increase in expression of TGF-β1 expression. Both of the above were compared to the WT group. Knockdown of IL-17A also decreased anoikis resistance in human TNBC and the murine 4T1 cell lines. Kaplan–Meier analysis disclosed a negative correlation between tumor expression of IL-17A and OS in TNBC patients.

Conclusion

We conclude that IL-17A promotes migratory and angiogenic activity in tumors, enhances anoikis resistance, and modulates the immune landscape of the tumor microenvironment such changes favor cancer metastasis.

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Data availability

All data generated or analysed during this study are included in this published article (and its Supplementary Information files).

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Acknowledgements

We are in debt to Chou, SS, Wang, YL, and Hsu, TH for their technique supports. This work was supported by the biobank from the Division of Experimental Surgery, Department of Surgery, Taipei Veterans General Hospital. We thank the Taiwan Animal Consortium (MOST 107-2319-B-001-002)--Taiwan Mouse Clinic, which is funded by the Ministry of Science and Technology (MOST) of Taiwan, for technical support in the IVIS animal experiment. We also thanks Ralph Kirby for English edition of this manuscript. Funding was provided by Ministry of Health and Welfare (Grant No. MOHW108-TDU-B-212-112015) and Health Promotion Administration, Ministry of Health and Welfare (Grant No. MOHW109-TDU-B-212-010001).

Funding

This work was supported by grants from the Ministry of Health and Welfare (Center of Excellence for Cancer Research at Taipei Veterans General Hospital phase II, MOHW108-TDU-B-212–112,015, phase III, MOHW109-TDU-B-212–010,001).

Author information

Author notes

  1. Ling-Ming Tseng, Jen-Hwey Chiu contributed equally in this manuscript.

Authors and Affiliations

  1. Comprehensive Breast Health Center & Division of General Surgery, Department of Surgery, Taipei Veterans General Hospital, No. 201, Sec. II, Shipai Rd, Taipei, 112, Taiwan, ROC

    Yi-Fang Tsai, Chi-Cheng Huang, Yen-Shu Lin, Chun-Yu Liu, Jen-Hwey Chiu & Ling-Ming Tseng

  2. Department of Surgery, School of Medicine, National Yang-Ming University, Taipei, Taiwan, ROC

    Yi-Fang Tsai & Ling-Ming Tseng

  3. Department of Public Health, College of Public Health, National Taiwan University, Taipei, Taiwan, ROC

    Chi-Cheng Huang

  4. Department of Pathology and Laboratory Medicine, Taipei Veterans General Hospital, Taipei, Taiwan, ROC

    Chih-Yi Hsu

  5. School of Medicine, National Yang-Ming University, Taipei, Taiwan, ROC

    Chih-Yi Hsu

  6. Faculty of Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan, ROC

    Ching-Po Huang

  7. Division of Medical Oncology, Department of Oncology, Taipei Veterans General Hospital, Taipei, Taiwan, ROC

    Chun-Yu Liu

  8. Department of Surgery, Cheng-Hsin General Hospital, Taipei, Taiwan, ROC

    Jen-Hwey Chiu

  9. Institute of Traditional Medicine, School of Medicine, National Yang-Ming University, Taipei, Taiwan, ROC

    Jen-Hwey Chiu

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  1. Yi-Fang Tsai
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Contributions

JHC formed the idea. LMT and JHC contributed equally in this manuscript. LMT supervised the experiments. CYL, LMT, YFT, CCH, and YSL provided clinical samples and data. CPH performed the experiments. CYH read the pathology slices.

Corresponding author

Correspondence to Jen-Hwey Chiu.

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Conflict of interest

We declare that we have no conflicts of interest, including financial and non-financial interests such as the following items.

  • Unpaid membership in a government or non-governmental organization.

  • Unpaid membership in an advocacy or lobbying organization.

  • Unpaid advisory position in a commercial organization.

  • Writing or consulting for an educational company.

  • Acting as an expert witness.

Ethics approval and consent to participate

Study protocols involving experimental mice followed ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines and were approved by the Institutional Animal Committee of and Taipei Veterans General Hospital (No. 2018–029). The human study for tumor tissue utilization from the biobank was approved by the Institutional Review Board of Taipei Veterans General Hospital (# 2013–10-020BC).

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Tsai, YF., Huang, CC., Lin, YS. et al. Interleukin 17A promotes cell migration, enhances anoikis resistance, and creates a microenvironment suitable for triple negative breast cancer tumor metastasis. Cancer Immunol Immunother 70, 2339–2351 (2021). https://doi.org/10.1007/s00262-021-02867-x

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