miR-17-5p suppresses cell proliferation and invasion by targeting ETV1 in triple-negative breast cancer.
miR-17-5p suppresses cell proliferation and invasion by targeting ETV1 in triple-negative breast cancer.
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DOI：
10.1186/s12885-017-3674-x
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发表时间：
2017-11-10
期刊：
影响因子：
3.8
通讯作者：
Wang W
中科院分区：
文献类型：
--
作者：
Li J;Lai Y;Ma J;Liu Y;Bi J;Zhang L;Chen L;Yao C;Lv W;Chang G;Wang S;Ouyang M;Wang W
三阴性乳腺癌（TNBC）是所有乳腺癌亚型中预后最差的恶性肿瘤。我们报道了ETV1在TNBC肿瘤发生中是一个重要的癌基因。因此，研究ETV1的关键调控microrna （mirna）可能有助于TNBC的靶向治疗。我们分别采用原位杂交（ISH）和免疫组织化学（IHC）检测TNBC患者样本中miR-17-5p和ETV1的位置。采用实时荧光定量PCR （qRT-PCR）检测miR-17-5p在TNBC组织和细胞系中的表达。采用qRT-PCR、western blotting和IHC检测ETV1的表达。利用细胞计数试剂盒-8 （CCK-8）、菌落形成、Transwell和伤口闭合试验来确定TNBC细胞的增殖和迁移能力。在斑马鱼模型中进行体内肿瘤转移试验。在TNBC细胞系和临床TNBC组织中，miR-17-5p的丰度显著降低。miR-17-5p表达水平与肿瘤大小（P < 0.05）、TNM分期（P < 0.05）密切相关。相比之下，ETV1在TNBC细胞系和组织中的表达明显上调。miR-17-5p的表达状态与ETV1呈负相关（r =−0.28,P = 3.88 × 10−3）。荧光素酶报告基因实验证实ETV1是miR-17-5p的直接靶点。在MDA-MB-231或BT549细胞中强制表达miR-17-5p可显著降低ETV1表达，抑制细胞体外增殖、迁移和体内肿瘤转移。然而，在miR-17-5p存在下挽救ETV1的表达可显著恢复细胞表型。无论是etv1阳性组还是etv1阴性组，miR-17-5p的高表达均与预后显著良好相关（log-rank检验，P < 0.001; P < 0.001）。单因素和多因素分析均显示miR-17-5p和ETV1是影响TNBC患者预后的独立危险因素。我们的数据表明，miR-17-5p通过靶向ETV1在TNBC中发挥肿瘤抑制作用，低丰度的miR-17-5p可能参与TNBC的发病机制。这些发现表明miR-17-5p可能是TNBC的治疗靶点。本文的在线版本（10.1186/s12885-017-3674-x）包含补充材料，授权用户可使用。
Triple-negative breast cancer (TNBC) is the malignancy with the worst outcome among all breast cancer subtypes. We reported that ETV1 is a significant oncogene in TNBC tumourigenesis. Consequently, investigating the critical regulatory microRNAs (miRNAs) of ETV1 may be beneficial for TNBC targeted therapy. We performed in situ hybridization (ISH) and immunohistochemistry (IHC) to detect the location of miR-17-5p and ETV1 in TNBC patient samples, respectively. miR-17-5p expression in TNBC tissues and cell lines was assessed by quantitative real-time PCR (qRT-PCR). ETV1 expression was evaluated by qRT-PCR, western blotting and IHC. Cell Counting Kit-8 (CCK-8), colony formation, Transwell and wound closure assays were utilized to determine the TNBC cell proliferation and migration capabilities. In vivo tumour metastatic assays were performed in a zebra fish model. The abundance of miR-17-5p was significantly decreased in TNBC cell lines and clinical TNBC tissues. The miR-17-5p expression levels were closely correlated with tumour size (P < 0.05) and TNM stage (P < 0.05). By contrast, the expression of ETV1 was significantly up-regulated in TNBC cell lines and tissues. There is an inverse correlation between the expression status of miR-17-5p and ETV1 (r = −0.28, P = 3.88 × 10−3). Luciferase reporter assay confirmed that ETV1 was a direct target of miR-17-5p. Forced expression of miR-17-5p in MDA-MB-231 or BT549 cells significantly decreased ETV1 expression and suppressed cell proliferation, migration in vitro and tumour metastasis in vivo. However, rescuing the expression of ETV1 in the presence of miR-17-5p significantly recovered the cell phenotype. High miR-17-5p expression was associated with a significantly favourable prognosis, in either the ETV1-positive or ETV1-negative groups (log-rank test, P < 0.001; P < 0.001). Both univariate and multivariate analyses showed that miR-17-5p and ETV1 were independent risk factors in the prognosis of TNBC patient. Our data indicate that miR-17-5p acts as a tumour suppressor in TNBC by targeting ETV1, and a low-abundance of miR-17-5p may be involved in the pathogenesis of TNBC. These findings indicate that miR-17-5p may be a therapeutic target for TNBC. The online version of this article (10.1186/s12885-017-3674-x) contains supplementary material, which is available to authorized users.
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影响因子：
--
作者：
Cohen R;Greenberg E;Nemlich Y;Schachter J;Markel G
通讯作者：
Markel G
影响因子：
8
作者：
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通讯作者：
Safe, S.
影响因子：
5.3
作者：
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通讯作者：
Saunders, Grady F.
影响因子：
4
作者：
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通讯作者：
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影响因子：
4.4
作者：
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通讯作者：
Wang, Wenjian