Inhibition of long non-coding RNA UCA1 by CRISPR/Cas9 attenuated malignant phenotypes of bladder cancer.
Inhibition of long non-coding RNA UCA1 by CRISPR/Cas9 attenuated malignant phenotypes of bladder cancer.
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CRISPR/Cas9抑制长链非编码RNA UCA1可减弱膀胱癌的恶性表型
DOI：
10.18632/oncotarget.14176
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发表时间：
2017-02-07
期刊：
影响因子：
--
通讯作者：
Li X
中科院分区：
文献类型：
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作者：
Zhen S;Hua L;Liu YH;Sun XM;Jiang MM;Chen W;Zhao L;Li X
CRISPR/Cas9是一种新型有效的基因组编辑技术，但其应用尚未广泛扩展到操纵长链非编码RNA（lncRNA）表达。尿路上皮癌相关的lncRNA 1（UCA 1）在膀胱癌中上调，并促进膀胱癌的进展。在这里，我们设计了对UCA 1特异性的gRNA，并构建了靶向UCA 1的CRISPR/Cas9系统。单一的CRISPR/Cas9-UCA 1转染5637和T24膀胱癌细胞可以有效抑制UCA 1的表达，而两种最有效的CRISPR/Cas9-UCA 1联合转染可以产生更满意的抑制效果。CRISPR/Cas9-UCA 1 s通过靶向基因组特异性DNA切割减弱UCA 1表达，从而在体外和体内显著抑制细胞增殖、迁移和侵袭。与CRISPR/Cas9-UCA 1对膀胱癌恶性表型的抑制作用相关的机制归因于诱导细胞周期停滞在G1期，细胞凋亡的大幅增加和MMPs活性的增强。此外，尿UCA 1可用作膀胱癌的非侵入性诊断标志物，如荟萃分析所示。总的来说，我们的数据表明，CRISPR/Cas9技术可用于下调lncRNA表达，尿UCA 1可用作诊断膀胱癌的非侵入性标志物。
CRISPR/Cas9 is a novel and effective genome editing technique, but its application is not widely expanded to manipulate long non-coding RNA (lncRNA) expression. The lncRNA urothelial carcinoma-associated 1 (UCA1) is upregulated in bladder cancer and promotes the progression of bladder cancer. Here, we design gRNAs specific to UCA1 and construct CRISPR/Cas9 systems targeting UCA1. Single CRISPR/Cas9-UCA1 can effectively inhibit UCA1 expression when transfected into 5637 and T24 bladder cancer cells, while the combined transfection of the two most effective CRISPR/Cas9-UCA1s can generate more satisfied inhibitory effect. CRISPR/Cas9-UCA1s attenuate UCA1 expression via targeted genome-specific DNA cleavage, resulting in the significant inhibition of cell proliferation, migration and invasion in vitro and in vivo. The mechanisms associated with the inhibitory effect of CRISPR/Cas9-UCA1 on malignant phenotypes of bladder cancer are attributed to the induction of cell cycle arrest at G1 phase, a substantial increase of apoptosis, and an enhanced activity of MMPs. Additionally, urinary UCA1 can be used as a non-invasive diagnostic marker for bladder cancer as revealed by a meta-analysis. Collectively, our data suggest that CRISPR/Cas9 technique can be used to down-modulate lncRNA expression, and urinary UCA1 may be used as a non-invasive marker for diagnosis of bladder cancer.