EZH2-mediated Epigenetic Silencing of miR-29/miR-30 targets LOXL4 and contributes to Tumorigenesis, Metastasis, and Immune Microenvironment Remodeling in Breast Cancer
EZH2-mediated Epigenetic Silencing of miR-29/miR-30 targets LOXL4 and contributes to Tumorigenesis, Metastasis, and Immune Microenvironment Remodeling in Breast Cancer
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EZH2 介导的 miR-29/miR-30 表观遗传沉默以 LOXL4 为靶标，有助于乳腺癌的肿瘤发生、转移和免疫微环境重塑
DOI：
10.7150/thno.44849
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发表时间：
2020-01-01
期刊：
影响因子：
12.4
通讯作者：
Zhang, Rui
中科院分区：
文献类型：
--
作者：
Yin, Huilong;Wang, Yidi;Zhang, Rui
Zeste增强子同源物2（EZH 2）是一种关键的表观遗传调节因子，参与乳腺癌的进展和转移。L0 XL 4越来越被认为是癌症进展中的重要参与者。迄今为止，EZH 2如何在乳腺癌进展中调节LOXL 4仍不清楚。研究方法：我们通过starBase v2.0分析评估了LOX家族蛋白与EZH 2在浸润性乳腺癌中的相关性，并使用Oncomine数据集评估了其与乳腺肿瘤发生的相关性。然后，我们将miRcode数据与基因表达综合数据（GEO）数据相结合，以筛选通过EZH 2介导LOXL 4调节的候选miRNA。我们通过qRT-PCR、蛋白质印迹、细胞增殖、集落形成和伤口愈合测定、异种移植实验、双荧光素酶报告基因测定和染色质免疫沉淀来探索EZH 2、miR-29 b/miR-30 d和LOXL 4在乳腺癌细胞中的调控机制。所有统计学检验均为双侧检验。结果如下：抑制EZH 2或LOXL 4或miR-29 b/miR-30 d过表达可降低体外和体内乳腺癌细胞增殖、迁移和转移。L0 XL 4被鉴定为miR-29 b和miR-30 d的直接靶标。EZH 2抑制通过启动子结合活性增强miR-30 d和miR-29 b转录，导致LOXL 4表达降低。人乳腺癌标本的免疫组织化学分析和小鼠肿瘤浸润巨噬细胞的流式细胞术分析显示EZH 2与LOXL 4表达和巨噬细胞浸润呈正相关。结论：我们的研究结果证实EZH 2-miR-29 b/miR-30 d-LOXL 4信号通路参与了乳腺癌的发生，并提示表观遗传调节是通过控制巨噬细胞活化来治疗乳腺癌的潜在靶点。
Enhancer of Zeste Homolog 2 (EZH2), a key epigenetic regulator, is involved in breast cancer progression and metastasis. LOXL4 is increasingly recognized as an important player in cancer progression. To date, how EZH2 regulates LOXL4 in the progression of breast cancer remains unclear. Methods: We evaluated the association between LOX family proteins and EZH2 in invasive breast carcinoma through the starBase v2.0 analysis, and its correlation with breast tumorigenesis using the Oncomine dataset. We then applied miRcode data combined with gene expression omnibus (GEO) data to screen candidate miRNAs mediating the regulation of LOXL4 by EZH2. We explored the regulatory mechanism of EZH2, miR-29b/miR-30d, and LOXL4 in breast cancer cells by qRT-PCR, Western blotting, cell proliferation, colony formation, and wound healing assays, xenograft experiments, dual-luciferase reporter assay, and chromatin immunoprecipitation. All statistical tests were two-sided. Results: Inhibition of EZH2 or LOXL4, or miR-29b/miR-30d overexpression, decreased breast cancer cell proliferation, migration, and metastasis in vitro and in vivo. LOXL4 was identified as a direct target of miR-29b and miR-30d. EZH2 inhibition enhanced miR-30d and miR-29b transcription via promoter binding activity, leading to the reduced expression of LOXL4. Immunohistochemical analysis of human breast cancer specimens and flow cytometry analysis of tumor-infiltrating macrophages in mice showed a positive association of EZH2 with LOXL4 expression and macrophage infiltration. Conclusions: Our findings identified EZH2-miR-29b/miR-30d-LOXL4 signaling pathway was involved in breast tumorigenesis, and suggested that the epigenetic modulation represents a potential therapeutic target for breast cancer by controlling macrophage activation.