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.
Oncotarget. 2016 Dec 25 [Epub ahead of print]
Shuai Zhen, Ling Hua, Yun-Hui Liu, Xiao-Min Sun, Meng-Meng Jiang, Wei Chen, Le Zhao, Xu Li
Center for Translational Medicine, The First Affiliated Hospital, Xi'an Jiaotong University, Xi'an 710061, P.R. China., Department of Veterinary Medicine, Rongchang Campus of Southwest University, Chongqing 402460, P.R. China., Department of Pharmacology and Toxicology, Beijing Institute of Radiation Medicine, Beijing 100850, P.R. China., Center for Laboratory Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, P.R. China., State Key Laboratory of Cancer Biology, Fourth Military Medical University, Xi'an 710032, Shaanxi, China.