title: Inhibition of long non-coding RNA UCA1 by CRISPR/Cas9 attenuated malignant phenotypes of bladder cancer.
abstract: 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.
authors.affiliation: Center for Translational Medicine, The First Affiliated Hospital, Xi'an Jiaotong University, Xi'an 710061, P.R. China.|Key Laboratory for Tumor Precision Medicine of Shaanxi Province, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, P.R. China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Zhen
authors.affiliation: Department of Veterinary Medicine, Rongchang Campus of Southwest University, Chongqing 402460, P.R. China.
authors.forename: Ling
authors.initials: L
authors.lastname: Hua
authors.affiliation: Department of Pharmacology and Toxicology, Beijing Institute of Radiation Medicine, Beijing 100850, P.R. China.
authors.forename: Yun-Hui
authors.initials: YH
authors.lastname: Liu
authors.affiliation: Center for Laboratory Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, P.R. China.
authors.forename: Xiao-Min
authors.initials: XM
authors.lastname: Sun
authors.affiliation: State Key Laboratory of Cancer Biology, The Fourth Military Medical University, Xi'an 710032, Shaanxi, China.
authors.forename: Meng-Meng
authors.initials: MM
authors.lastname: Jiang
authors.affiliation: Center for Laboratory Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, P.R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.affiliation: Center for Translational Medicine, The First Affiliated Hospital, Xi'an Jiaotong University, Xi'an 710061, P.R. China.|Key Laboratory for Tumor Precision Medicine of Shaanxi Province, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, P.R. China.
authors.forename: Le
authors.initials: L
authors.lastname: Zhao
authors.affiliation: Center for Translational Medicine, The First Affiliated Hospital, Xi'an Jiaotong University, Xi'an 710061, P.R. China.|Key Laboratory for Tumor Precision Medicine of Shaanxi Province, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, P.R. China.
authors.forename: Xu
authors.initials: X
authors.lastname: Li
mesh_terms: D000818:Animals; D017209:Apoptosis; D014408:Biomarkers, Tumor / Q000235:genetics* / Q000652:urine; D064130:CRISPR-Associated Proteins / Q000235:genetics* / Q000378:metabolism; D064113:CRISPR-Cas Systems; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D015536:Down-Regulation; D059585:G1 Phase Cell Cycle Checkpoints; D015972:Gene Expression Regulation, Neoplastic; D055785:Gene Knockdown Techniques; D018390:Gene Targeting / Q000379:methods*; D006801:Humans; D008297:Male; D020782:Matrix Metalloproteinases / Q000378:metabolism; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009361:Neoplasm Invasiveness; D010641:Phenotype; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics / Q000378:metabolism; D062085:RNA, Long Noncoding / Q000235:genetics* / Q000652:urine; D013997:Time Factors; D014162:Transfection; D047368:Tumor Burden; D001749:Urinary Bladder Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology
chemical_list: D014408:Biomarkers, Tumor; D064130:CRISPR-Associated Proteins; D000094704:RNA, Guide, CRISPR-Cas Systems; D062085:RNA, Long Noncoding; C515159:UCA1 RNA, human; D020782:Matrix Metalloproteinases
keywords: CRISPR/Cas9; UCA1; bladder cancer; long non-coding RNA