Synthetic Artificial Long Non-coding RNA Shows Higher Efficiency in Specific Malignant Phenotype Inhibition Compared to the CRISPR/Cas Systems.
Synthetic Artificial Long Non-coding RNA Shows Higher Efficiency in Specific Malignant Phenotype Inhibition Compared to the CRISPR/Cas Systems.
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与 CRISPR/Cas 系统相比，合成的人工长非编码 RNA 在特定恶性表型抑制方面表现出更高的效率
DOI：
10.3389/fmolb.2020.617600
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发表时间：
2020
影响因子：
5
通讯作者：
Zhang Q
中科院分区：
文献类型：
--
作者：
Yao L;Zhang Q;Li A;Ma B;Zhang Z;Liu J;Liang L;Zhu S;Gan Y;Zhang Q
目的：致癌转录因子和微小RNA分别通过转录和转录后调节在人类癌症中发挥重要作用。这些现象提出了关于人工装置同时调节miRNA和TF的能力的问题。在这项研究中，我们的目的是构建一个人工长的非编码RNA，“alncRNA”，它模仿CRISPR/Cas系统，并阐明其在膀胱癌细胞系中的治疗作用。同时，我们还比较了alncRNA和CRISPR/Cas系统在调控基因表达方面的效率。研究设计和方法：根据合成生物学的工程原理，我们将转录因子适体的串联cDNA序列与miRNA结合位点的串联cDNA拷贝相结合，构建了alncRNA。为了证明该平台的实用性，我们选择β -catenin、NF-κB、miR-940和miR-495作为功能靶标，并使用膀胱癌细胞系5637和T24作为测试模型。采用实时荧光定量PCR（qPCR）、双荧光素酶法和相关表型实验分别检测相关基因的表达和治疗效果。结果：双荧光素酶法检测alncRNA对转录因子的转录活性有抑制作用。qPCR结果显示，alncRNA能降低相关TF靶基因和miRNA的表达水平，抑制效果优于CRIPSR dCas 9-KRAB。通过功能实验，在alncRNA感染的膀胱细胞中观察到细胞增殖减少，凋亡增加和运动抑制。结论：总之，我们的合成装置确实起到抗肿瘤调节剂的作用，其在膀胱癌细胞中同步完成转录和转录后调节，并且与CRISPR/Cas系统相比，在特定恶性表型抑制方面显示出更高的效率。最重要的是，合成的alncRNA在膀胱癌细胞系中诱导了抗癌作用。因此，我们的设备为癌症治疗提供了一种新的策略，并可能成为癌细胞的有用“武器”。
Objective: Both oncogenic transcription factors (TFs) and microRNAs (miRNAs) play an important regulator in human cancer by transcriptional and post-transcriptional regulation, respectively. These phenomena raise questions about the ability of artificial device to regulate miRNAs and TFs simultaneously. In this study, we aimed to construct an artificial long non-coding RNA, “alncRNA,” which imitated CRISPR/Cas systems and to illuminate its therapeutic effects in bladder cancer cell lines. At the same time, we also compared the efficiency of alncRNA and CRISPR/Cas systems in regulating gene expression. Study Design and Methods: Based on engineering principles of synthetic biology, we combined tandem arrayed cDNA sequences of aptamer for TFs with tandem arrayed cDNA copies of binding sites for the miRNAs to construct alncRNA. In order to prove the utility of this platform, we chose β -catenin, NF-κB, miR-940, and miR-495 as the functional targets and used the bladder cancer cell lines 5637 and T24 as the test models. Real-time Quantitative PCR (qPCR), dual-luciferase assay and relative phenotypic experiments were applied to severally test the expression of relative gene and therapeutic effects of our devices. Result: Dual-luciferase assay indicated alncRNA could inhibit transcriptional activity of TFs. What’s more, the result of qPCR showed that expression levels of the relative TFs target genes and miRNAs were reduced by corresponding alncRNA and the inhibitory effect was better than CRIPSR dCas9-KRAB. By functional experiments, decreased cell proliferation, increased apoptosis, and motility inhibition were observed in alncRNA-infected bladder cells. Conclusion: In summary, our synthetic devices indeed function as anti-tumor regulator, which synchronously accomplish transcriptional and post-transcriptional regulation in bladder cancer cell and show higher efficiency in specific malignant phenotype inhibition compared to the CRISPR/Cas systems. Most importantly, Anti-cancer effects were induced by the synthetic alncRNA in the bladder cancer lines. Our devices, therefore, provides a novel strategy for cancer therapy and could be a useful “weapon” for cancer cell.
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影响因子：
3.1
作者：
Wang R;Wu Y;Huang W;Chen W
通讯作者：
Chen W
影响因子：
--
作者：
Liu, Yuchen;Huang, Weiren;Cai, Zhiming
通讯作者：
Cai, Zhiming
Huachansu通过FAS/FASL和TNF-Alpha/TNFR1途径在体外和体内抑制人膀胱癌细胞的生长。
DOI：
10.1186/s13046-015-0134-9
发表时间：
2015-02-25
期刊：
Journal of experimental & clinical cancer research : CR
影响因子：
--
作者：
Yang T;Shi R;Chang L;Tang K;Chen K;Yu G;Tian Y;Guo Y;He W;Song X;Xu H;Ye Z
通讯作者：
Ye Z
DOI：
10.1038/s41580-018-0028-8
发表时间：
2018-10
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
Haberle V;Stark A
通讯作者：
Stark A
强力霉素通过PAR-1/NF-κB/miR-17/E-cadherin通路抑制乳腺癌EMT和转移
影响因子：
--
作者：
Zhong W;Chen S;Qin Y;Zhang H;Wang H;Meng J;Huai L;Zhang Q;Yin T;Lei Y;Han J;He L;Sun B;Liu H;Liu Y;Zhou H;Sun T;Yang C
通讯作者：
Yang C