Taraxacum mongolicum extract inhibited malignant phenotype of triple-negative breast cancer cells in tumor-associated macrophages microenvironment through suppressing IL-10/STAT3/PD-L1 signaling pathways
Taraxacum mongolicum extract inhibited malignant phenotype of triple-negative breast cancer cells in tumor-associated macrophages microenvironment through suppressing IL-10/STAT3/PD-L1 signaling pathways
复制标题
蒲公英提取物通过抑制IL-10/STAT3/PD-L1信号通路抑制肿瘤相关巨噬细胞微环境中三阴性乳腺癌细胞的恶性表型
DOI：
10.1016/j.jep.2021.113978
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发表时间：
2021-03-23
影响因子：
5.4
通讯作者：
Han, Shu-Yan
中科院分区：
文献类型：
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作者：
Deng, Xin-Xin;Jiao, Yan-Na;Han, Shu-Yan
民族药理学相关性：三阴性乳腺癌（TNBC）是最具侵袭性和预后最差的乳腺癌，治疗选择有限。蒲公英（Taraxacum mongolicum）是一种传统中药，自古以来就被用于治疗乳腺炎、乳腺脓肿和乳腺增生。在现代药理学研究中，已证实其具有抗乳腺癌活性。我们以前报道过黄芪提取物可以诱导TNBC细胞凋亡。然而，其在肿瘤微环境中的抗肿瘤作用和机制尚未阐明。研究目的：肿瘤相关巨噬细胞（TAMs）在调节肿瘤细胞与免疫系统之间的相互作用中发挥重要作用。本研究旨在探讨黄芪提取物在TAM微环境下对TNBC细胞的作用及其机制，以及对M2巨噬细胞极化的影响。材料与方法：M2巨噬细胞经佛波醇-12-肉豆蔻酸酯13-乙酸酯（PMA）和白细胞介素4（IL-4）诱导，并通过流式细胞术、定量RT-PCR（qRT-PCR）、Western印迹和ELISA进行验证。将MDA-MB-468 TNBC细胞与M2巨噬细胞的上清液共培养，其提供TAMs微环境。采用MTT法检测甘草提取物对TNBC细胞的抗肿瘤活性。通过transwell测定法测量TNBC细胞的侵袭和迁移能力。Western blotting和qRT-PCR分别检测蛋白和基因的表达。结果：TAMs微环境促进TNBC细胞的增殖、迁移和侵袭。但黄芪提取物可抑制TAMs诱导的MDA-MB-231和MDA-MB-468细胞的恶性转化。TAMs和IL-10均能诱导TNBC细胞中STAT 3活化和PD-L1高表达，而IL-10中和抗体可减弱这种作用。丹参提取物在TAMs微环境下对TNBC细胞中的STAT 3和PD-L1具有抑制作用。此外，在M2巨噬细胞中，黄芪提取物显著促进M1样标志物TNF-α、IL-8和iNOS的表达，但降低M2样标志物IL-10、CD 206、精氨酸酶-1和TGF-β的表达。结论：丹参提取物通过抑制IL-10/STAT 3/PD-L1免疫抑制信号通路抑制TNBC细胞在TAMs微环境中的增殖、迁移和侵袭。此外，黄芪提取物促进了巨噬细胞从M2表型向M1表型的极化。因此，我们的研究结果表明，通过调节肿瘤免疫微环境，TNBC可能是一种有前途的治疗策略。
Ethnopharmacological relevance: Triple-negative breast cancer (TNBC) is the most aggressive and the worst prognosis breast cancer with limited treatment options. Taraxacum mongolicum (also called dandelion) is a traditional Chinese medicine has been used to treat mastitis, breast abscess, and hyperplasia of mammary glands since ancient times. In modern pharmacological research, dandelion has been proven with anti-breast cancer activities. We previously reported that dandelion extract could induce apoptosis in TNBC cells. However, its antitumor effects and mechanisms in the tumor microenvironment have not yet been elucidated. Aim of the study: Tumor-associated macrophages (TAMs) play an important role in regulating the interaction between tumor cells and the immune system. The present study aimed to investigate the effects and mechanisms of dandelion extract on TNBC cells under the microenvironment of TAMs, as well as its influence on the polarization of M2 macrophages. Materials and methods: M2 macrophages were induced by phorbol-12-myristate 13-acetate (PMA) and interleukin 4 (IL-4), and verified by flow cytometry, quantitative RT-PCR (qRT-PCR), Western blotting, and ELISA. MDA-MB231 and MDA-MB-468 TNBC cells were co-cultured with the supernatant of M2 macrophage which providing the TAMs microenvironment. The antitumor activity of dandelion extract in TNBC cells was evaluated by MTT assay. The invasive and migratory capacity of TNBC cells was measured by transwell assays. The expression of protein and gene was assessed by Western blotting and qRT-PCR, respectively. Results: TAMs microenvironment promoted the proliferation, migration, and invasion of TNBC cells. However, dandelion extract inhibited the malignant property of MDA-MB-231 and MDA-MB-468 cells induced by TAMs. Both of TAMs and IL-10 caused STAT3 activation and PD-L1 higher expression, the immunosuppressive molecules in TNBC cells, and this effect can be attenuated by IL-10 neutralizing antibody. Dandelion extract exerted inhibition on STAT3 and PD-L1 in TNBC cells under TAMs microenvironment. Furthermore, in M2 macrophages, dandelion extract remarkably promoted the expression of M1-like marker TNF-alpha, IL-8, and iNOS, but reduced M2-like marker IL-10, CD206, Arginase-1, and TGF-beta. Conclusion: Dandelion extract inhibited the proliferation, migration and invasion of TNBC cells in TAMs microenvironment through suppressing IL-10/STAT3/PD-L1 immunosuppressive signaling pathway. Furthermore, dandelion extract promoted the polarization of macrophages from M2 to M1 phenotype. Thus, our results indicated that dandelion may serve as a promising therapeutic strategy for TNBC by modulating tumor immune microenvironment.