MiRNA-Embedded ShRNAs for Radiation-Inducible LGMN Knockdown and the Antitumor Effects on Breast Cancer.
MiRNA-Embedded ShRNAs for Radiation-Inducible LGMN Knockdown and the Antitumor Effects on Breast Cancer.
复制标题
嵌入 miRNA 的 ShRNA 用于辐射诱导 LGMN 敲低以及对乳腺癌的抗肿瘤作用
DOI：
10.1371/journal.pone.0163446
复制
发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Wang XY
中科院分区：
文献类型：
--
作者：
Zhang ZQ;Cao Z;Liu C;Li R;Wang WD;Wang XY
Legumain（LGMN）在乳腺癌（BC）和其他实体瘤中高度表达，是一个潜在的抗癌靶点。在这里，我们研究了短发夹RNA（shRNAs）靶向嵌入microRNA-155（miR-155）架构中的LGMN的抗肿瘤作用，该架构由辐射诱导的嵌合RNA聚合酶II（Pol II）启动子驱动。用嵌合启动子产生慢病毒载体，所述嵌合启动子控制下游shRNA-miR-155盒的表达。通过使用共聚焦显微镜观察荧光。采用实时定量PCR和Western blotting检测LGMN、MMP 2和MMP 9的表达水平。通过平板集落形成实验和侵袭实验分析BC细胞的增殖和侵袭能力。在这里，我们证明了嵌合启动子可以有效地诱导辐射处理。此外，靶向LGMN的shRNA-miR-155盒可以被嵌合启动子有效激活。辐射加LGMN的敲低损害BC细胞中的集落形成并抑制细胞迁移和侵袭。抑制LGMN下调BC细胞中MMP 2和MMP 9的表达。Pol II驱动的shRNA-miR-155能有效抑制BC细胞的生长和侵袭能力，且这种干扰作用受辐射剂量的调节。此外，LGMN的敲低通过调节MMPs的表达来增强BC细胞的侵袭性表型。
Legumain (LGMN) is highly expressed in breast cancer (BC) and other solid tumors and is a potential anticancer target. Here we investigate the anti-tumor effects of short hairpin RNAs (shRNAs) targeting LGMN embedded in a microRNA-155 (miR-155) architecture, which is driven by a radiation-inducible chimeric RNA polymerase II (Pol II) promoter. Lentiviral vectors were generated with the chimeric promoter which controlled the expression of downstream shRNA-miR-155 cassette. Fluorescence was observed by using confocal microscopy. Real-time quantitative PCR and Western blotting were used to determine the expression level of LGMN, MMP2, and MMP9. Furthermore, the proliferation and invasive ability of BC cells was analyzed via plate colony formation and invasion assays. Here we demonstrated that the chimeric promoter could be effectively induced by radiation treatment. Furthermore, the shRNA-miR-155 cassette targeting LGMN could be effectively activated by the chimeric promoter. Radiation plus knockdown of LGMN impairs colony formation and dampens cell migration and invasion in BC cells. Inhibition of LGMN downregulates MMP2 and MMP9 expression in BC cells. Pol II-driven shRNA-miR-155 could effectively suppress the growth and invasiveness of BC cells, and that the interference effects could be regulated by radiation doses. Moreover, knockdown of LGMN alleviates the aggressive phenotype of BC cells through modulating MMPs expression.
登录
查看更多内容
影响因子：
82.9
作者：
通讯作者：
--
影响因子：
--
作者：
Wu, Mei;Shao, Guang-Rui;Ruan, Zheng-Min
通讯作者：
Ruan, Zheng-Min
影响因子：
64.8
作者：
Grimm, Dirk;Streetz, Konrad L.;Kay, Mark A.
通讯作者：
Kay, Mark A.
影响因子：
8.4
作者：
Haugen, Mads H.;Boye, Kjetil;Flatmark, Kjersti
通讯作者：
Flatmark, Kjersti
影响因子：
14.9
作者：
Fowler DK;Williams C;Gerritsen AT;Washbourne P
通讯作者：
Washbourne P