Synthetic miRNA-mowers targeting miR-183-96-182 cluster or miR-210 inhibit growth and migration and induce apoptosis in bladder cancer cells.
Synthetic miRNA-mowers targeting miR-183-96-182 cluster or miR-210 inhibit growth and migration and induce apoptosis in bladder cancer cells.
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靶向 miR-183-96-182 簇或 miR-210 的合成 miRNA 割草机抑制膀胱癌细胞的生长和迁移并诱导细胞凋亡。
DOI：
10.1371/journal.pone.0052280
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Cai Z
中科院分区：
文献类型：
--
作者：
Liu Y;Han Y;Zhang H;Nie L;Jiang Z;Fa P;Gui Y;Cai Z
microRNAs（miRNAs）是人类肿瘤生物学行为的内源性调节因子。据报道，几种天然非编码RNA通过碱基配对相互作用抑制miRNA。这些现象提出了关于人工装置调节miRNA的能力的问题。本研究的目的是创建针对单个miRNA或miRNA簇的合成装置，并确定它们对膀胱癌细胞表型的治疗效果。在SV-40启动子驱动的海肾荧光素酶基因的3′非翻译区（UTR）插入串联的凸起的miRNA结合位点，构建两个抑制miR-183-96-182簇或miR-210的“miRNA-mowers”。产生具有不与任何已知miRNA互补的串联重复序列的第三装置作为非靶向对照。在功能分析中，膀胱癌T24和UM-UC-3细胞用三种装置中的每一种转染，然后进行检测其影响的测定。荧光素酶测定表明，在miRNA-割草机中的荧光素酶报告基因的活性降低至未靶向对照的30-50%。使用实时qPCR，靶miRNA的表达水平显示通过相应的miRNA-mower降低2-3倍。MTT法、流式细胞术和体外划痕实验分别检测细胞生长、凋亡和迁移。细胞生长抑制，增加凋亡，并减少运动中观察到的miRNA-割草机转染的膀胱癌细胞。这种模块化设计策略不仅可以阻断单个靶miRNA，而且可以阻断靶miRNA簇的整个成员。在膀胱癌细胞系中通过合成的miRNA-切割器诱导抗癌作用。miR-183/96/182簇和miR-210在膀胱癌中发挥致癌作用。本研究为miRNA功能缺失研究和肿瘤治疗提供了一个潜在的合成生物学平台。
MicroRNAs (miRNAs) function as endogenous regulators of biological behaviors of human cancers. Several natural non-coding RNAs are reported to inhibit miRNAs by base-pairing interactions. These phenomena raise questions about the ability of artificial device to regulate miRNAs. The purpose of this study is to create synthetic devices that target a single miRNA or a miRNA cluster and to ascertain their therapeutic effects on the phenotypes of bladder cancer cells. Tandem bulged miRNA binding sites were inserted into the 3′ untranslated region (UTR) of the SV-40 promoter-driven Renilla luciferase gene to construct two “miRNA-mowers” for suppression of miR-183-96-182 cluster or miR-210. A third device with tandem repeat sequences not complementary to any known miRNA was generated as an untargeted-control. In functional analyses, bladder cancer T24 and UM-UC-3 cells were transfected with each of the three devices, followed by assays for detection of their impacts. Luciferase assays indicated that the activities of the luciferase reporters in the miRNA-mowers were decreased to 30–50% of the untargeted-control. Using Real-Time qPCR, the expression levels of the target miRNAs were shown to be reduced 2-3-fold by the corresponding miRNA-mower. Cell growth, apoptosis, and migration were tested by MTT assay, flow cytometry assay, and in vitro scratch assay, respectively. Cell growth inhibition, increased apoptosis, and decreased motility were observed in miRNA-mowers-transfected bladder cancer cells. Not only a single target miRNA but also the whole members of a target miRNA cluster can be blocked using this modular design strategy. Anti-cancer effects are induced by the synthetic miRNA-mowers in the bladder cancer cell lines. miR-183/96/182 cluster and miR-210 are shown to play oncogenic roles in bladder cancer. A potentially useful synthetic biology platform for miRNA loss-of-function study and cancer treatment has been established in this work.
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影响因子：
3.7
作者：
Nardinocchi L;Pantisano V;Puca R;Porru M;Aiello A;Grasselli A;Leonetti C;Safran M;Rechavi G;Givol D;Farsetti A;D'Orazi G
通讯作者：
D'Orazi G
影响因子：
2.8
作者：
Carradori, Simone;Cristini, Cristiano;Di Pierro, Giovanni Battista
通讯作者：
Di Pierro, Giovanni Battista
影响因子：
4.8
作者：
Kim, Ha Won;Haider, Husnain K.;Ashraf, Muhammad
通讯作者：
Ashraf, Muhammad
DOI：
10.1126/science.1187197
发表时间：
2010-06-18
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Cazalla D;Yario T;Steitz JA
通讯作者：
Steitz JA
影响因子：
14.8
作者：
Liang, Chun-Chi;Park, Ann Y.;Guan, Jun-Lin
通讯作者：
Guan, Jun-Lin