Exosome derived from epigallocatechin gallate treated breast cancer cells suppresses tumor growth by inhibiting tumor-associated macrophage infiltration and M2 polarization.
Exosome derived from epigallocatechin gallate treated breast cancer cells suppresses tumor growth by inhibiting tumor-associated macrophage infiltration and M2 polarization.
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DOI：
10.1186/1471-2407-13-421
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发表时间：
2013-09-17
期刊：
影响因子：
3.8
通讯作者：
Kim CW
中科院分区：
文献类型：
--
作者：
Jang JY;Lee JK;Jeon YK;Kim CW
肿瘤相关巨噬细胞（TAM）在肿瘤微环境中发挥着重要作用。特别是，M2 巨噬细胞有助于肿瘤进展，具体取决于 NF-κB 的表达。肿瘤来源的外泌体可以通过将 miRNA 转移到免疫细胞来调节肿瘤微环境。表没食子儿茶素没食子酸酯 (EGCG) 具有众所周知的抗肿瘤作用；然而，没有关于 EGCG 对与癌细胞和 TAM 通讯的影响的数据。小鼠乳腺癌细胞系 4T1 用于体内和离体研究。从EGCG处理的4T1细胞中提取外泌体，并使用微阵列筛选miRNA的变化。从小鼠肿瘤移植物中分离出的肿瘤细胞或 TAM 与源自 EGCG 处理和/或 miR-16 抑制剂转染的 4T1 细胞的外泌体一起孵育。通过 RT-qPCR 或蛋白质印迹评估单核细胞趋化因子（CSF-1 和 CCL-2）、M2 巨噬细胞中高表达（IL-6 和 TGF-β）和低表达（TNF-α）的细胞因子以及 NF-κB 通路中的分子（IKKα 和 Iκ-B）。 EGCG 抑制小鼠乳腺癌模型中的肿瘤生长，这与 TAM 和 M2 巨噬细胞浸润减少有关。在 EGCG 处理的小鼠肿瘤细胞中，单核细胞趋化因子（CSF-1 和 CCL-2）的表达较低，并且 EGCG 导致 TAM 细胞因子从 M2- 转变为 M1 样表型，IL-6 和 TGF-β 减少以及 TNF-α 增加证明了这一点。将分离的肿瘤细胞与 EGCG 离体孵育可抑制 CSF-1 和 CCL-2 的表达。 TAM 与 EGCG 处理的 4T1 细胞的外泌体进行离体孵育，导致 IKKα 抑制并伴随 I-κB 积累； IL-6和TGF-β增加；以及TNF-α的减少。 EGCG 上调 4T1 细胞和外泌体中的 miR-16。用来自 EGCG 处理和 miR-16 敲低的 4T1 细胞的外泌体处理肿瘤细胞或 TAM，恢复了 EGCG 处理的外泌体对趋化因子、细胞因子和 NF-κB 通路的上述影响。我们的数据表明，EGCG 上调肿瘤细胞中的 miR-16，miR-16 可通过外泌体转移至 TAM，并抑制 TAM 浸润和 M2 极化。我们提出了一种新机制，即 EGCG 通过调节肿瘤微环境中的 TAM 发挥抗肿瘤活性。
Tumor-associated macrophages (TAM) play an important role in tumor microenvironment. Particularly, M2 macrophages contribute to tumor progression, depending on the expression of NF-κB. Tumor-derived exosomes can modulate tumor microenvironment by transferring miRNAs to immune cells. Epigallocatechin gallate (EGCG) has well known anti-tumor effects; however, no data are available on the influence of EGCG on communication with cancer cells and TAM. Murine breast cancer cell lines, 4T1, was used for in vivo and ex vivo studies. Exosome was extracted from EGCG-treated 4T1 cells, and the change of miRNAs was screened using microarray. Tumor cells or TAM isolated from murine tumor graft were incubated with exosomes derived from EGCG-treated and/or miR-16 inhibitor-transfected 4T1 cells. Chemokines for monocytes (CSF-1 and CCL-2), cytokines both with high (IL-6 and TGF-β) and low (TNF-α) expression in M2 macrophages, and molecules in NF-κB pathway (IKKα and Iκ-B) were evaluated by RT-qPCR or western blot. EGCG suppressed tumor growth in murine breast cancer model, which was associated with decreased TAM and M2 macrophage infiltration. Expression of chemokine for monocytes (CSF-1 and CCL-2) were low in tumor cells from EGCG-treated mice, and cytokines of TAM was skewed from M2- into M1-like phenotype by EGCG as evidenced by decreased IL-6 and TGF-β and increased TNF-α. Ex vivo incubation of isolated tumor cells with EGCG inhibited the CSF-1 and CCL-2 expression. Ex vivo incubation of TAM with exosomes from EGCG-treated 4T1 cells led to IKKα suppression and concomitant I-κB accumulation; increase of IL-6 and TGF-β; and, decrease of TNF-α. EGCG up-regulated miR-16 in 4T1 cells and in the exosomes. Treatment of tumor cells or TAM with exosomes derived from EGCG-treated and miR-16-knock-downed 4T1 cells restored the above effects on chemokines, cytokines, and NF-κB pathway elicited by EGCG-treated exosomes. Our data demonstrate that EGCG up-regulates miR-16 in tumor cells, which can be transferred to TAM via exosomes and inhibits TAM infiltration and M2 polarization. We suggest a novel mechanism by which EGCG exerts anti-tumor activity via regulation of TAM in tumor microenvironment.
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