XIAOPI formula inhibits the pre-metastatic niche formation in breast cancer via suppressing TAMs/CXCL1 signaling
XIAOPI formula inhibits the pre-metastatic niche formation in breast cancer via suppressing TAMs/CXCL1 signaling
复制标题
小皮方通过抑制TAMs/CXCL1信号传导抑制乳腺癌转移前生态位形成
DOI：
10.1186/s12964-020-0520-6
复制
发表时间：
2020-03-26
影响因子：
8.4
通讯作者：
Wang, Zhiyu
中科院分区：
文献类型：
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作者：
Zheng, Yifeng;Wang, Neng;Wang, Zhiyu
研究背景近年来的研究表明，转移前小生境（PMN）是介导肿瘤转移的先决条件。本课题前期研究表明消痞方通过抑制肿瘤相关巨噬细胞（TAMs）分泌的CXCL 1，从而抑制乳腺癌肺转移。本研究旨在探讨消痞方对乳腺癌中性粒细胞（PMN）形成的影响及其分子机制。采用CCK 8和EdU染色法检测消痞方对TAMs和4 T1细胞在共培养体系中增殖的影响。采用Transwell实验检测消痞方对HSPCs和4 T1. 0细胞侵袭能力的影响。通过将乳腺癌T1细胞移植到Balb/c小鼠的乳腺垫中，并通过荧光素酶成像监测肺转移。免疫荧光法检测肺组织上皮-间质转化过程及中性粒细胞（PMN）的形成。采用流式细胞仪检测消痞方对TAMs表型、造血干/祖细胞（HSPCs）和髓源性抑制细胞（MDSCs）的影响。而对M1型巨噬细胞无明显影响。随后发现TAM/CXCL 1信号转导刺激c-Kit+/Sca-1+ HSPC的募集及其分化为CD 11b +/Gr-1+ MDSC，这是解释PMN形成的象征性事件。此外，消痞方还能有效抑制HSPCs的活化，抑制HSPCs与TAMs共培养体系诱导的乳腺癌细胞4 T1的增殖和转移，提示消痞方在体外具有抑制PMN生成的作用。乳腺癌异种移植实验进一步证明消痞方在体内可抑制乳腺癌PMN的形成及随后的肺转移。消痞方治疗后骨髓HSPCs和肺组织MDSCs数量均明显减少。然而，TAMs中CXCL 1的过表达可减轻消痞方的抑制作用。结论综上所述，本研究为消痞方预防乳腺癌PMN形成提供了临床前证据，并强调TAMs/CXCL 1是PMN靶向治疗的潜在治疗策略。
BackgroundRecent findings suggested that premetastatic niche (PMN) is a prerequisite in mediating cancer metastasis. Previously we demonstrated that XIAOPI formula could inhibit breast cancer lung metastasis via inhibiting tumor associated macrophages (TAMs)-secreted CXCL1. Herein, we aimed to explore the effects of XIAOPI formula on preventing breast cancer PMN formation and its underlying molecular mechanisms.MethodsCXCL1 expression of TAMs was detected by qPCR and Western blotting assay. The influences of XIAOPI formula on the proliferation of TAMs and 4 T1 in the co-culture system were tested by CCK8 or EdU staining. Transwell experiment was applied to determine the effects of XIAOPI formula on the invasion ability of HSPCs and 4 T1. Breast cancer xenografts were built by inoculating 4 T1 cells into the mammary pads of Balb/c mice and lung metastasis was monitored by luciferase imaging. Immune fluorescence assay was used to test the epithelial-mesenchymal transition process and PMN formation in the lung tissues. The effects of XIAOPI formula on TAMs phenotype, hematopoietic stem/progenitor cells (HSPCs) and myeloid-derived suppressor cells (MDSCs) were determined by flow cytometry.ResultsIt was found that XIAOPI formula could inhibit the proliferation and polarization of M2 phenotype macrophages, and reduce CXCL1 expression in a dose-dependent manner. However, M1 phenotype macrophages were not significantly affected by XIAOPI formula. TAMs/CXCL1 signaling was subsequently found to stimulate the recruitment of c-Kit+/Sca-1+HSPCs and their differentiation into CD11b+/Gr-1+MDSCs, which were symbolic events accounting for PMN formation. Moreover, XIAOPI formula was effective in inhibiting HSPCs activation and suppressing the proliferation and metastasis of breast cancer cells 4 T1 induced by HSPCs and TAMs co-culture system, implying that XIAOPI was effective in preventing PMN formation in vitro. Breast cancer xenograft experiments further demonstrated that XIAOPI formula could inhibit breast cancer PMN formation and subsequent lung metastasis in vivo. The populations of HSPCs in the bone marrow and MDSCs in the lung tissues were all remarkably declined by XIAOPI formula treatment. However, the inhibitory effects of XIAOPI formula could be relieved by CXCL1 overexpression in the TAMs.ConclusionsTaken together, our study provided preclinical evidence supporting the application of XIAOPI formula in preventing breast cancer PMN formation, and highlighted TAMs/CXCL1 as a potential therapeutic strategy for PMN targeting therapy.Video AbstractGraphical abstract