Synthetic Circular RNA Functions as a miR-21 Sponge to Suppress Gastric Carcinoma Cell Proliferation.
Synthetic Circular RNA Functions as a miR-21 Sponge to Suppress Gastric Carcinoma Cell Proliferation.
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DOI：
10.1016/j.omtn.2018.09.010
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发表时间：
2018-12-07
期刊：
影响因子：
--
通讯作者：
Meltzer SJ
中科院分区：
文献类型：
--
作者：
Liu X;Abraham JM;Cheng Y;Wang Z;Wang Z;Zhang G;Ashktorab H;Smoot DT;Cole RN;Boronina TN;DeVine LR;Talbot CC Jr;Liu Z;Meltzer SJ
含有miR结合序列的microRNA（mir）构成了一种潜在的强大分子治疗策略，天然存在的圆形RNA（CIRCRNA）被证明是癌细胞中有效的mir海绵。针对特定miR的功能。去磷酸化和磷酸化后，使用T4 RNA连接酶1使用5'-3'终端结合来实现循环。使用RNase R和胎儿牛血清评估CircRNA稳定性。在三种胃癌细胞中，荧光素酶报告者，细胞增殖和细胞凋亡测定也对下游蛋白的影响也被串联描绘质量标签（TMT）标记（通过蛋白酶换识识别剂骄傲：PXD008584），然后是蛋白质印迹，我们包括对抗核酸酶消化的人工circrna海绵。 miR-21在包括癌蛋白DAXX在内的下游蛋白质靶标的活性。简单，有效，方便的策略，以实现体外MIR功能的靶向丧失，并在人类患者中潜在的未来治疗应用。
MicroRNA (miR) sponges containing miR binding sequences constitute a potentially powerful molecular therapeutic strategy. Recently, naturally occurring circular RNAs (circRNAs) were shown to function as efficient miR sponges in cancer cells. We hypothesized that synthetic circRNA sponges could achieve therapeutic loss-of-function targeted against specific miRs. Linear RNA molecules containing miR-21 binding sites were transcribed in vitro; after dephosphorylation and phosphorylation, circularization was achieved using 5′-3′ end-ligation by T4 RNA ligase 1. circRNA stability was assessed using RNase R and fetal bovine serum. Competitive inhibition of miR-21 activity by a synthetic circRNA sponge was assessed using luciferase reporter, cell proliferation, and cell apoptosis assays in three gastric cancer cell lines. circRNA effects on downstream proteins were also delineated by Tandem Mass Tag (TMT) labeling (data available via ProteomeXchange identifier PRIDE: PXD008584), followed by western blotting. We conclude that artificial circRNA sponges resistant to nuclease digestion can be synthesized using simple enzymatic ligation steps. These sponges inhibit cancer cell proliferation and suppress the activity of miR-21 on downstream protein targets, including the cancer protein DAXX. In summary, synthetic circRNA sponges represent a simple, effective, convenient strategy for achieving targeted loss of miR function in vitro, with potential future therapeutic application in human patients.
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影响因子：
48
作者：
Gentner, Bernhard;Schira, Giulia;Naldini, Luigi
通讯作者：
Naldini, Luigi
影响因子：
21.3
作者：
Ma, Li;Young, Jennifer;Prabhala, Harsha;Pan, Elizabeth;Mestdagh, Pieter;Muth, Daniel;Teruya-Feldstein, Julie;Reinhardt, Ferenc;Onder, Tamer T.;Valastyan, Scott;Westermann, Frank;Speleman, Frank;Vandesompele, Jo;Weinberg, Robert A.
通讯作者：
Weinberg, Robert A.
影响因子：
14.9
作者：
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通讯作者：
Futreal PA
影响因子：
7.3
作者：
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通讯作者：
Tuschl, Thomas
DOI：
10.1016/j.jcmgh.2017.02.003
发表时间：
2017-05
影响因子：
7.2
作者：
Liu X;Meltzer SJ
通讯作者：
Meltzer SJ