Tetrastigma polysaccharide reprogramming of tumor-associated macrophages via PPARγ signaling pathway to play antitumor activity in breast cancer
Tetrastigma polysaccharide reprogramming of tumor-associated macrophages via PPARγ signaling pathway to play antitumor activity in breast cancer
复制标题
DOI：
10.1016/j.jep.2023.116645
复制
发表时间：
2023-05-22
影响因子：
5.4
通讯作者：
Fu, Huiying
中科院分区：
文献类型：
--
作者：
Liu, Xia;Liu, Xianli;Fu, Huiying
民族药理意义：红花蛇舌草是中国民间传说中用于抗肿瘤治疗的一种典型的畲族药物。SYQ多糖(SYQ-PA)具有抗氧化、抗炎等作用，但其抗肿瘤作用及其机制尚不清楚。本研究的目的：探讨SYQ-PA的体内外抗乳腺癌作用及其机制。材料和方法：本研究以4周龄和8周龄的MMTV-PYMT小鼠为模型，研究SYQ-PA在体内乳腺癌发生中的作用。用IL4/13诱导的小鼠腹腔巨噬细胞模型探讨其作用机制。采用流式细胞术分析肿瘤微环境和巨噬细胞分型的变化。用xCELLigence系统检测巨噬细胞条件培养液对乳腺癌细胞的抑制作用。用细胞微球阵列检测炎症因子。采用共培养系统检测细胞的迁移和侵袭情况。结果：SYQ-PA可显著抑制MMTV-PyMT小鼠乳腺原发肿瘤的生长过程，减少TAMs的侵袭，促进M1表型极化。体外研究表明，SYQ-PA能促进巨噬细胞从IL4/13诱导的M2向抗肿瘤的M1表型极化，诱导的巨噬细胞的条件培养液(CM)抑制乳腺癌细胞的增殖。同时，SYQ-PA处理的巨噬细胞在共培养体系中可抑制4T1细胞的迁移和侵袭。进一步的结果表明，SYQ-PA抑制抗炎因子的释放，促进炎性细胞因子的产生，从而可能诱导M1巨噬细胞极化，抑制乳腺癌细胞的增殖。随后，基于RNAseq和分子分析的潜在机制分析表明，SYQ-PA抑制巨噬细胞中PPARγ的表达，并调节其下游的核因子-kappaB。经PPARγ抑制剂T0070907处理后，SYQ-PA的作用减弱，甚至消失。结论：SYQ-PA对乳腺癌的抑制作用至少部分是通过PPARγ激活和β-catenin介导的M2巨噬细胞极化来实现的。这些数据阐明了SYQ-PA的抗肿瘤作用及其机制，为SYQ-PA作为乳腺癌巨噬细胞瘤免疫治疗的辅助药物提供了可能。
Ethnopharmacological relevance: Tetrastigma Hemsleyanum Diels et Gilg (SYQ) is a typical She ethnomedicine that has been used in anti-tumor treatment in Chinese folklore. The polysaccharide of SYQ (SYQ-PA) has been reported to have antioxidant and anti-inflammatory effects, but the effect and mechanism on antitumor is still unclear. Aim of the study: To investigate the activity and mechanism of SYQ-PA against breast cancer in vitro and in vivo. Materials and methods: In this study, different stages of MMTV-PYMT mice, which at 4-week-old and 8-week-old representative the transition from hyperplasia to late carcinoma, were used to investigate the potential effect of SYQ-PA of breast cancer development in vivo. The mechanism was explored with IL4/13-induced peritoneal macrophages model. Flow cytometry assay was employed to analysis the change of tumor microenvironment and the macrophages typing. The inhibition of the condition medium from macrophages on breast cancer cells was detected with xCELLigence system detection. The inflammation factors were tested with cytometric bead array. Co-culture system was used to detect the cell migration and invasion. In addition, the underlying mechanism was investigated using RNAseq analysis, Q-PCR and Western blot, and the PPAR gamma inhibitor was used to verify the mechanism.Results: SYQ-PA significantly attenuated the process of breast primary tumor growth and reduced the infiltration of TAMs accompanied promoting the polarization of M1 phenotype in MMTV-PyMT mice. Then in vitro studies showed that SYQ-PA promoted macrophages polarization form IL4/13 induced M2 toward to the anti-tumor M1 phenotypes, and the conditioned medium (CM) from the induced macrophages inhibited the proliferation of breast cancer cells. At the same time, SYQ-PA treated macrophages inhibited the migration and invasion of 4T1 in the co-culture system. Further results indicated that SYQ-PA suppressed the release of anti-inflammatory factors and promoted the production of inflammatory cytokines which may induce M1 macrophage polariza-tion and inhibit breast cancer cell proliferation. Subsequently, the underlying mechanism analysis based on RNAseq and molecular assays indicated that SYQ-PA inhibited PPAR gamma expression and regulated downstream NF-kappa B in macrophages. After treated with PPAR gamma inhibitor, T0070907, the effect of SYQ-PA was decreased, or even disappeared. As the downstream, the expression of beta-catenin was also inhibited obviously, those above all contribute the process of SYQ-PA induced M1 macrophages polarization.Conclusions: Collectively, SYQ-PA was observed inhibited breast cancer, at least in part, via PPAR gamma activation-and beta-catenin-mediated M2 macrophages polarization. These data expound the antitumor effect and mechanism of SYQ-PA, and provide a possible that SYQ-PA can be used as an adjuvant drug for macrophage tumor immunotherapy in breast cancer.