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Studying the Effects of Tumor-Secreted Paracrine Ligands on Macrophage Activation using Co-Culture with Permeable Membrane Supports
Published on: November 28, 2019
Cancer-derived exosomal miR-138-5p modulates polarization of tumor-associated macrophages through inhibition of KDM6B
Jing Xun1,2, Lingfang Du1, Ruifang Gao1
1School of Medicine, Nankai University, Tianjin 300071, China.
Abstract:
Rationale: Differential activation of macrophages correlates closely with tumor progression, and the epigenetic factor lysine demethylase 6B (KDM6B, previously named JMJD3) mediates the regulation of macrophage polarization through an unknown mechanism. Methods: We developed a suspension coculture system comprising breast cancer cells and macrophages and used RT-qPCR and western blotting to measure KDM6B expression. Bioinformatics and luciferase reporter assays were used to identify candidate microRNAs of cancer cells responsible for the downregulation of KDM6B. To determine if exosomes mediated the transfer of miR-138-5p between cancer cells to macrophages, we treated macrophages with exosomes collected from the conditioned medium of cancer cells. The effects of exosomal miR-138-5p on macrophage polarization were measured using RT-qPCR, flow cytometry, and chromatin immunoprecipitation assays. We employed a mouse model of breast cancer, metastatic to the lung, to evaluate the effects on tumor metastasis of macrophages treated with miR-138-5p-enriched exosomes. To develop a diagnostic evaluation index, the levels of exosomal miR-138-5p in samples from patients with breast cancer were compared to those of controls. Results: Coculture of breast cancer cells led to downregulation of KDM6B expression in macrophages. Cancer cell-derived exosomal miR-138-5p inhibited M1 polarization and promoted M2 polarization through inhibition of KDM6B expression in macrophages. Macrophages treated with exosomal miR-138-5p promoted lung metastasis, and the level of circulating exosomal miR-138-5p positively correlated with the progression of breast cancer. Conclusion: Our data suggest that miR-138-5p was delivered from breast cancer cells to tumor-associated macrophages via exosomes to downregulate KDM6B expression, inhibit M1 polarization, and stimulate M2 polarization. Therefore, exosomal miR-138-5p represents a promising prognostic marker and target for the treatment of breast cancer.
Insights
Breast cancer cells release exosomal miR-138-5p, which inhibits KDM6B in macrophages, promoting M2 polarization and metastasis. This exosomal microRNA is a potential biomarker for breast cancer progression and a therapeutic target.
Area of Science:
- Molecular Biology
- Cancer Research
- Immunology
Background:
- Macrophage polarization is critical in tumor progression.
- Lysine demethylase 6B (KDM6B) regulates macrophage polarization via an unknown mechanism.
- Epigenetic regulation in cancer-associated macrophages is an area of intense research.
Purpose of the Study:
- To elucidate the mechanism by which breast cancer cells influence macrophage polarization.
- To investigate the role of KDM6B in cancer-associated macrophage polarization.
- To identify potential diagnostic markers and therapeutic targets for breast cancer.
Main Methods:
- Developed a co-culture system of breast cancer cells and macrophages.
- Utilized RT-qPCR, western blotting, bioinformatics, and luciferase assays to identify miR-138-5p and its target KDM6B.
- Investigated exosome-mediated transfer of miR-138-5p and its effects on macrophage polarization and tumor metastasis in vitro and in vivo mouse models.
Main Results:
- Breast cancer cell-derived exosomal miR-138-5p downregulates KDM6B expression in macrophages.
- Exosomal miR-138-5p promotes M2 macrophage polarization and inhibits M1 polarization.
- Macrophages treated with exosomal miR-138-5p enhance lung metastasis in a mouse model; circulating exosomal miR-138-5p levels correlate with breast cancer progression.
Conclusions:
- Exosomal miR-138-5p from breast cancer cells targets KDM6B in tumor-associated macrophages, driving M2 polarization and promoting metastasis.
- Exosomal miR-138-5p serves as a promising prognostic biomarker for breast cancer.
- Targeting exosomal miR-138-5p offers a potential therapeutic strategy for breast cancer treatment.
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