miR-145-dependent targeting of Junctional Adhesion Molecule A and modulation of fascin expression are associated with reduced breast cancer cell motility and invasiveness
miR-145-dependent targeting of Junctional Adhesion Molecule A and modulation of fascin expression are associated with reduced breast cancer cell motility and invasiveness
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DOI：
10.1038/onc.2010.386
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发表时间：
2010-12-01
期刊：
影响因子：
8
通讯作者：
Yip, G. W.
中科院分区：
文献类型：
--
作者：
Goette, M.;Mohr, C.;Yip, G. W.
microRNA是一类小的非编码RNA，在转录和翻译水平上调节细胞和发育的基本过程。在乳腺癌中，与健康对照组织相比，miR-145表达下调。由于miR-145的几个预测靶点可能调节细胞运动性，我们旨在研究miR-145在乳腺癌细胞运动性和侵袭性中的潜在作用。在Affyssin阵列技术的辅助下，我们证明了在MDA-MB-231、MCF-7、MDA-MB-468和SK-BR-3乳腺癌细胞以及石川子宫内膜癌细胞中miR-145的过表达导致细胞-细胞粘附蛋白JAM-A和肌动蛋白捆绑蛋白fascin的下调。此外，podocalyxin和Serpin E1 mRNA水平下调，γ-肌动蛋白，transgelin和MYL9上调后，miR-145过表达。这些miR-145依赖性表达变化显著降低了癌细胞的运动性，如通过延时视频显微镜、划痕伤口闭合测定和基质胶侵袭测定所揭示的。免疫荧光显微镜显示肌动蛋白细胞骨架的重组和细胞形态的变化，miR-145过表达，导致更多的皮质肌动蛋白分布，减少肌动蛋白应力纤维和丝状伪足的形成。在10%的pre-miR-145转染的MDA-MB-231细胞中观察到核旋转，伴随着核周肌动蛋白的减少。荧光素酶活化试验证实了JAM-A的3'UTR的直接miR-145依赖性调节，而siRNA介导的JAM-A表达的敲低导致MDA-MB-231和MCF-7乳腺癌细胞的运动性和侵袭性降低。我们的数据将JAM-A和肌成束蛋白确定为miR-145的新靶点，牢固地确立了miR-145在调节乳腺癌细胞运动性中的作用。我们的数据为未来的miR-145靶向抗转移性癌症治疗方法提供了理论基础。Oncogene（2010）29，6569 - 6580; doi：10.1038/onc.2010.386; 2010年9月6日在线发表
Micro RNAs are small non-coding RNAs, which regulate fundamental cellular and developmental processes at the transcriptional and translational level. In breast cancer, miR-145 expression is downregulated compared with healthy control tissue. As several predicted targets of miR-145 potentially regulate cell motility, we aimed at investigating a potential role for miR-145 in breast cancer cell motility and invasiveness. Assisted by Affymetrix array technology, we demonstrate that overexpression of miR-145 in MDA-MB-231, MCF-7, MDA-MB-468 and SK-BR-3 breast cancer cells and in Ishikawa endometrial carcinoma cells leads to a downregulation of the cell-cell adhesion protein JAM-A and of the actin bundling protein fascin. Moreover, podocalyxin and Serpin E1 mRNA levels were downregulated, and gamma-actin, transgelin and MYL9 were upregulated upon miR-145 overexpression. These miR-145-dependent expression changes drastically decreased cancer cell motility, as revealed by time-lapse video microscopy, scratch wound closure assays and matrigel invasion assays. Immunofluorescence microscopy demonstrated restructuring of the actin cytoskeleton and a change in cell morphology by miR-145 overexpression, resulting in a more cortical actin distribution, and reduced actin stress fiber and filopodia formation. Nuclear rotation was observed in 10% of the pre-miR-145 transfected MDA-MB-231 cells, accompanied by a reduction of perinuclear actin. Luciferase activation assays confirmed direct miR-145-dependent regulation of the 3'UTR of JAM-A, whereas siRNA-mediated knockdown of JAM-A expression resulted in decreased motility and invasiveness of MDA-MB-231 and MCF-7 breast cancer cells. Our data identify JAM-A and fascin as novel targets of miR-145, firmly establishing a role for miR-145 in modulating breast cancer cell motility. Our data provide a rationale for future miR-145-targeted approaches of antimetastatic cancer therapy. Oncogene (2010) 29, 6569-6580; doi:10.1038/onc.2010.386; published online 6 September 2010