Inhibition of RAC1-GEF DOCK3 by miR-512-3p contributes to suppression of metastasis in non-small cell lung cancer
Inhibition of RAC1-GEF DOCK3 by miR-512-3p contributes to suppression of metastasis in non-small cell lung cancer
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DOI：
10.1016/j.biocel.2015.02.005
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发表时间：
2015-04-01
影响因子：
4
通讯作者：
Zhou, Caicun
中科院分区：
文献类型：
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作者：
Zhu, Xingli;Gao, Guanghui;Zhou, Caicun
MicroRNA是一类调控基因表达的非编码小RNA。在这项研究中，我们证明了维甲酸（RA）处理增加了miR-512- 3 p的表达。miR-512- 3 p过表达抑制非小细胞肺癌（NSCLC）细胞系A549和H1299中的细胞粘附、迁移和侵袭。miR-512- 3 p抑制剂在稳定表达miR-512的H1299细胞中部分逆转了这些作用。我们鉴定了DOCK 3（一种RAC 1-GEF（guanine nucleotide exchange factor））作为miR-512- 3 p的靶基因。过表达miR-512- 3 p可导致DOCK 3蛋白表达减少，但其mRNA表达无明显变化。DOCK 3的敲低导致与miR-512- 3 p过表达所观察到的粘附、迁移和侵袭类似的作用。RAC 1 pull-down活性检测结果表明，过表达miR-512- 3 p可显著降低RAC 1活性，且降低效率高于DOCK 3敲低。此外，与配对的正常对照相比，miR-512- 3 p在大多数NSCLC患者肿瘤样品中的表达受到抑制，表明miR-512- 3 p可能在肺癌发展中起关键作用。总之，我们的结果支持miR-512- 3 p可以通过DOCK 3调节NSCLC A549和H1299细胞系中RAC 1的活性来抑制肿瘤细胞的粘附、迁移和侵袭。(C)2015爱思唯尔有限公司版权所有。
MicroRNAs are a class of small non-coding RNAs regulating gene expression. In this study, we demonstrated that retinoic acid (RA) treatment increases the expression of miR-512-3p. Overexpression of miR-512-3p inhibited cell adhesion, migration, and invasion in non-small cell lung cancer (NSCLC) cell lines A549 and H1299. miR-512-3p inhibitor partially reversed these effects in H1299 cells stably expressing miR-512. We identified DOCK3, a RAC1-GEF (guanine nucleotide exchange factor), as a target gene of miR-512-3p. Overexpression of miR-512-3p led to the decrease of DOCK3 protein but not its mRNA. Knockdown of DOCK3 resulted in similar effects on adhesion, migration, and invasion as observed of miR-512-3p overexpression. Active RAC1 pull-down assay indicated that overexpression of miR-512-3p could decrease the activity of RAC1 with a higher efficiency than that of DOCK3 knockdown. Furthermore, expression of miR-512-3p was suppressed in most NSCLC patient tumor samples compared to its paired normal controls, suggesting that miR-512-3p might play a crucial role in lung cancer development. In conclusion, our results supported that miR-512-3p could inhibit tumor cell adhesion, migration, and invasion by regulating the RAC1 activity via DOCK3 in NSCLC A549 and H1299 cell lines. (C) 2015 Elsevier Ltd. All rights reserved.