Downregulation of miRNA-141 in breast cancer cells is associated with cell migration and invasion: involvement of ANP32E targeting.
Downregulation of miRNA-141 in breast cancer cells is associated with cell migration and invasion: involvement of ANP32E targeting.
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DOI：
10.1002/cam4.1024
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发表时间：
2017-03
期刊：
影响因子：
4
通讯作者：
Zhu K
中科院分区：
文献类型：
--
作者：
Li P;Xu T;Zhou X;Liao L;Pang G;Luo W;Han L;Zhang J;Luo X;Xie X;Zhu K
MicroRNA (miRNA) 调节许多细胞活动，包括癌症的发生、进展和转移。一些miRNA参与乳腺癌（BC）的迁移和侵袭，从而影响患者的预后。进行微阵列分析以比较 BC 组织中的 miRNA 表达，并通过 qPCR 确认结果。使用微孔板 Transwell 测定在体外使用 MDA-MB-231 细胞研究 BC 细胞迁移和侵袭。在过度表达 miRNA 模拟物的细胞中，使用荧光素酶测定、qPCR 和蛋白质印迹分析来研究 miRNA 靶向。使用靶标 siRNA 慢病毒感染进行 miRNA 靶标的敲低。结果显示，与匹配的周围组织相比，乳腺癌肿瘤组织中的 microRNA-141 (miR-141) 表达下调。 miR-141 表达的下调与肿瘤分期、淋巴结受累以及 PCNA、Ki67 和 HER2 的表达相关。 miR-141 的过表达抑制 BC 细胞的体外增殖、迁移和侵袭。根据计算机分析结果，选择 ANP32E 基因作为进一步研究的假定靶标之一。双荧光素酶报告系统的结果表明 ANP32E 是 miR-141 的直接靶点。 miR-141 的过表达下调了 BC 细胞中 mRNA 和蛋白质水平的 ANP32E 表达。 ANP32E 的敲低可在体外抑制 BC 细胞的增殖、迁移和侵袭，类似于 miR-141 过表达的效果。我们的研究揭示了 miR-141 在 BC 生长和转移中发挥的重要作用。此外，我们首次将 ANP32E 确定为 miR-141 靶标之一，并证明其参与细胞增殖、迁移和侵袭的调节。
MicroRNAs (miRNAs) regulate many cellular activities, including cancer development, progression, and metastasis. Some miRNAs are involved in breast cancer (BC) migration and invasion, thus affect patients’ prognosis. Microarray analysis was performed to compare miRNA expression in BC tissues, and results confirmed by qPCR. BC cell migration and invasion were studied in vitro with MDA‐MB‐231 cells using microplate transwell assays. miRNA targeting was investigated using luciferase assays, qPCR, and Western blot analysis in cells with overexpression of miRNA mimics. Knockdown of miRNA targets was performed using target siRNA lentiviral infection. Results show that microRNA‐141 (miR‐141) was downregulated in breast cancer tumor tissues compared with matched surrounding tissues. Downregulation of miR‐141 expression correlated with tumor stage, lymph node involvement, and expressions of PCNA, Ki67, and HER2. Overexpression of miR‐141 inhibited BC cell proliferation, migration, and invasion in vitro. ANP32E gene was selected as one putative target for further studies based on results from in silico analysis. Results from a dual‐luciferase reporter system suggested ANP32E as a direct target of miR‐141. Overexpression of miR‐141 downregulated ANP32E expression at both mRNA and protein levels in BC cells. Knockdown of ANP32E inhibited BC cell proliferation, migration, and invasion in vitro, mimicking the effect of the overexpression of miR‐141. Our study revealed important roles miR‐141 plays in BC growth and metastasis. Moreover, for the first time, we identified ANP32E as one of the miR‐141 targets, and demonstrated its involvement in the regulation of cell proliferation, migration, and invasion.