Macrophage-mediated tumorigenic effects of breast cancer cell exosomes

Year 2023, Volume: 48 Issue: 1, 289 - 296, 31.03.2023

Pervin Elvan Tokgün , Samet Türel , Kubilay Incı , Büşra Çelikkaya , Onur Tokgün

Abstract

Purpose: As a heterogeneous disease, breast cancer is one of the most commonly diagnosed cancers due to hereditary/genetic factors and non-hereditary factors and is a leading cause of disease-associated death among women all over the world. Macrophages are one of the cell types that have essential roles in physiological and pathological cellular processes containing hemostasis, inflammation, and carcinogenesis. Exosomes secreted by cancer cells regulate cellular processes such as the polarization of macrophages, different organ metastasis, and drug resistance via internalized macrophages. In this study, we aimed to reveal the exosome-mediated regulation of the tumor microenvironment in specific to macrophage cells.
Materials and Methods: THP-1 monocyte cells are cultured with RPMI-1640 medium supplemented with 20 ng/ul phorbol-12-myristate-13-acetate (PMA) for 48 hours to induce macrophage differentiation. Exosomes of MCF-10A and MDA-MB-231 cells were isolated using a commercial kit. Fold changes in proliferation capacities of MCF-10A treated with exosomes of MDA-MB-231 cells were calculated.
Results: A statistically significant increase in the expression of genes involved in the inflammation-related JAK/STAT pathway in THP-1 cells treated with MDA-MB-231 exosomes was observed. Besides, we detected morphological and proliferative changes in MCF-10A cells that were co-cultured with THP-1/MDA-MB-231exo cells.
Conclusion: This study may contribute to the literature by showing that phenotypic differences in immune system cells can occur through exosomes in the tumor microenvironment.

Keywords

Macrophage differentiation, exosomes, breast cancer, inflammation

Meme kanseri hücre ekzozomlarının makrofaj aracılı tümörijenik etkileri

Abstract

Amaç: Kalıtsal ve genetik faktörler ile kalıtsal olmayan faktörler nedeniyle heterojen bir hastalık olan meme kanseri, tüm dünyada kadınlar arasında en sık görülen kanserlerden biridir ve hastalığa bağlı ölümlerin önde gelen nedenlerinden biridir. Makrofajlar hemostaz, inflamasyon ve karsinogenezi içeren fizyolojik ve patolojik hücresel süreçlerde önemli rollere sahip hücre tiplerinden biridir. Kanser hücreleri tarafından salgılanan eksozomlar, makrofajların polarizasyonu, farklı organ metastazı ve ilaç direnci gibi hücresel süreçleri düzenleyebilirler. Bu çalışmada makrofaj hücrelerine özgü tümör mikro çevresinin eksozom aracılı regülasyonunu ortaya koymayı amaçladık.
Gereç ve Yöntem: THP-1 monosit hücreleri, makrofaj farklılaşmasını indüklemek için 48 saat boyunca 20 ng/ul forbol-12-miristat-13-asetat (PMA) içeren RPMI-1640 besiyeri ile kültüre edildi. MCF-10A ve MDA-MB-231 hücrelerinden ticari bir kit kullanılarak eksozom izolasyonu gerçekleştirildi. MDA-MB-231 hücrelerinin eksozomları ile muamele edilen MCF-10A hücrelerinin proliferasyon kapasitelerindeki kat değişimleri değerlendirildi.
Bulgular: MDA-MB-231 eksozomları ile muamele edilen THP-1 hücrelerinde inflamasyon ile ilişkili JAK/STAT yolağında görev alan genlerin ifadesinde istatistiksel olarak anlamlı bir artış saptandı. Ayrıca THP-1/MDA-MB-231exo hücreleri ile birlikte kültüre edilen MCF-10A hücrelerinde morfolojik ve proliferatif değişiklikler saptandı.
Sonuç: Bu çalışmada tümör mikro çevresindeki eksozomlar aracılığıyla immün sistem hücrelerinde fenotipik farklılıkların oluşabileceğini göstererek literatüre katkı sağlayabilecektir.

Keywords

Makrofaj farklılaşması, eksozomlar, meme kanseri, inflamasyon

References

• Veronesi U, Boyle P, Goldhirsch A, Orecchia R, Viale G. Breast cancer. Lancet. 2005;365:1727-41.
• Guo W, Li Y, Pang W, Shen H. Exosomes: a potential therapeutic tool targeting communications between tumor cells and macrophages. Mol Ther. 2020;28:1953-64.
• Jabbari N, Akbariazar E, Feqhhi M, Rahbarghazi R, Rezaie J. Breast cancer-derived exosomes: tumor progression and therapeutic agents. J Cell Physiol. 2020;235:6345-56.
• Halvaei S, Daryani S, Eslami-S Z, Samadi T, Jafarbeik-Iravani N, Bakhshayesh TO et al. Exosomes in cancer liquid biopsy: a focus on breast cancer. Mol Ther Nucleic Acids. 2018;10:131-41.
• Tan Y, Luo X, Lv W, Hu W, Zhao C, Xiong M et al. Tumor-derived exosomal components: the multifaceted roles and mechanisms in breast cancer metastasis. Cell Death Dis. 2021;12:547.
• Yang E, Wang X, Gong Z, Yu M, Wu H, Zhang D. Exosome-mediated metabolic reprogramming: the emerging role in tumor microenvironment remodeling and its influence on cancer progression. Signal Transduct Target Ther. 2020;5:242.
• Yang SS, Ma S, Dou H, Liu F, Zhang SY, Jiang C et al. Breast cancer-derived exosomes regulate cell ınvasion and metastasis in breast cancer via mir-146a to activate cancer associated fibroblasts in tumor microenvironment. Exp Cell Res. 2020;391:111983.
• Chen Q, Li Y, Gao W, Chen L, Xu W, Zhu X. Exosome-mediated crosstalk between tumor and tumor-associated macrophages. Front Mol Biosci. 2021;8:764222.
• Lee CC, Lin JC, Hwang WL, Kuo YJ, Chen HK, Tai SK et al. Macrophage-secreted ınterleukin-35 regulates cancer cell plasticity to facilitate metastatic colonization. Nat Commun. 2018;9:3763.
• Liu Q, Zhao E, Geng B, Gao S, Yu H, He X et al. Tumor-associated macrophage-derived exosomes transmitting mir-193a-5p promote the progression of renal cell carcinoma via Timp2-dependent vasculogenic mimicry. Cell Death Dis. 2022;13:382.
• Othman N, Jamal R, Abu N. Cancer-derived exosomes as effectors of key inflammation-related players. Front Immunol. 2019;10:2103.
• DeNardo DG, Ruffell B. Macrophages as regulators of tumour immunity and immunotherapy. Nat Rev Immunol. 2019;19:369-82.
• Wu L, Zhang X, Zhang B, Shi H, Yuan X, Sun Y et al. Exosomes derived from gastric cancer cells activate nf-kb pathway in macrophages to promote cancer progression. Tumour Biol. 2016;37:12169–80.
• Chen G, Huang AC, Zhang W, Zhang G, Wu M, Xu W et al. Exosomal PD-L1 contributes to immunosuppression and is associated with anti-PD-1 response. Nature 2018;560:382–86.
• Zhou J, Li X, Wu X, Zhang T, Zhu Q, Wang X et al. Exosomes released from tumor-associated macrophages transfer miRNAs that induce a Treg/Th17 cell imbalance in epithelial ovarian cancer. Cancer Immunol Res. 2018;6:1578-92.
• Tokgün, O, Inci, K. Myc manipulates the miRNA content and biologic functions of small cell lung cancer cell-derived small extracellular vesicles. Mol Biol Rep. 2022;49:7953-65.
• Mildner A, Yona S, Jung S. A close encounter of the third kind: monocyte-derived cells. Adv Immunol. 2013;120:69-103.
• Nahrendorf M, Swirski FK, Aikawa E, Stangenberg L, Wurdinger T, Figueiredo JL et al. The healing myocardium sequentially mobilizes two monocyte subsets with divergent and complementary functions. J Exp Med. 2007;204:3037-47.
• Chomarat P, Banchereau J, Davoust J, Karolina Palucka A. IL-6 switches the differentiation of monocytes from dendritic cells to macrophages. Nat Immunol. 2020;1:510–14.
• Ushach I, Zlotnik A. Biological role of granulocyte macrophage colony-stimulating factor (GM-CSF) and macrophage colony-stimulating factor (M-CSF) on cells of the myeloid lineage. J Leukoc Biol. 2016;100:481–89.
• Hamilton JA. Colony-stimulating factors in inflammation and autoimmunity. Nat Rev Immunol. 2008;8:533–44.
• Sánchez-Martín L, Estecha A, Samaniego R, Sánchez-Ramón S, Vega MÁ, Sánchez-Mateos P. The chemokine CXCL12 regulates monocyte-macrophage differentiation and RUNX3 expression. Blood. 2011;117:88-97.
• Chanput W, Mes JJ, Wichers HJ. THP-1 cell line: an in vitro cell model for immune modulation approach. Int Immunopharmacol. 2014;23:37-45.
• Lund ME, To J, O'Brien BA, Donnelly S. The choice of phorbol 12-myristate 13-acetate differentiation protocol influences the response of THP-1 macrophages to a pro-inflammatory stimulus. J Immunol Methods. 2016;430:64-70.
• Zhou J, Tang Z, Gao S, Li C, Feng Y, Zhou X. Tumor-associated macrophages: recent insights and therapies. Front Oncol. 2020;10:188.
• Myers KV, Amend SR, Pienta KJ. Targeting Tyro3, Axl and MerTK (TAM receptors): implications for macrophages in the tumor microenvironment. Cancer. 2019;18:94.
• Chen Q, Li Y, Gao W, Chen L, Xu W, Zhu X. Exosome-mediated crosstalk between tumor and tumor-associated macrophages. Front Mol Biosci. 2021;8:764222.
• Baig MS, Roy A, Rajpoot S, Liu D, Savai R, Banerjee S et al. Tumor-derived exosomes in the regulation of macrophage polarization. Inflamm Res. 2020;69:435-51.
• Malla RR, Kiran P. Tumor microenvironment pathways: cross regulation in breast cancer metastasis. Genes Dis. 2020;9:310-24.
• Qiu P, Guo Q, Yao Q, Chen J, Lin J. Characterization of exosome-related gene risk model to evaluate the tumor ımmune microenvironment and predict prognosis in triple-negative breast cancer. Front Immunol. 2021;12:736030.
• Piombino C, Mastrolia I, Omarini C, Candini O, Dominici M, Piacentini F et al. The role of exosomes in breast cancer diagnosis. Biomedicines. 2021;9:312.
• Chaudhary P, Gibbs L, Maji S, Lewis C, Suzuki S, Vishwanatha J. Serum exosomal-annexin A2 is associated with African-American triple-negative breast cancer and promotes angiogenesis. Breast Cancer Res BCR. 2020;22:11.
• Lan J, Sun L, Xu F, Liu L, Hu F, Song D et al. M2 macrophage-derived exosomes promote cell migration and invasion in colon cancer. Cancer Res. 2019;79:146-58.
• Kalra H, Drummen GP, Mathivanan S. Focus on extracellular vesicles: Introducing the next small big thing. Int J Mol Sci. 2016;17:170.
• Xin L, Wu Y, Liu C, Zeng F, Wang JL, Wu DZ et al. Exosome-mediated transfer of lncRNA HCG18 promotes M2 macrophage polarization in gastric cancer. Mol Immunol. 2021;140:196-205.
• Zhao S, Mi Y, Guan B, Zheng B, Wei P, Gu Y et al. Tumor-derived exosomal miR-934 induces macrophage M2 polarization to promote liver metastasis of colorectal cancer. J Hematol Oncol. 2020;13:156.
• Pan Z, Zhao R, Li B, Qi Y, Qiu W, Guo Q et al. EWSR1-induced circNEIL3 promotes glioma progression and exosome-mediated macrophage immunosuppressive polarization via stabilizing IGF2BP3. Mol Cancer. 2022;21:16.
• Wan S, Zhao E, Kryczek I, Vatan L, Sadovskaya A, Ludema G et al. Tumor-associated macrophages produce interleukin 6 and signal via STAT3 to promote expansion of human hepatocellular carcinoma stem cells. Gastroenterology. 2014;147:1393–404.
• Ham S, Lima LG, Chai EPZ, Muller A, Lobb RJ, Krumeich S et al. Breast cancer-derived exosomes alter macrophage polarization via gp130/STAT3 signaling. Front Immunol. 2018;9:871.