PMID- 29254226
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20191120
IS  - 1949-2553 (Electronic)
IS  - 1949-2553 (Linking)
VI  - 8
IP  - 60
DP  - 2017 Nov 24
TI  - CRISPR-ON-Mediated KLF4 overexpression inhibits the proliferation, migration and 
      invasion of urothelial bladder cancer in vitro and in vivo.
PG  - 102078-102087
LID - 10.18632/oncotarget.22158 [doi]
AB  - Kruppel like factor 4 (KLF4), a transcription factor associated with 
      carcinogenesis and tumor progression, plays an important role in various 
      malignancies. In the present study, we utilized the CRISPR-ON system to 
      upregulate KLF4 expression level and subsequently investigated the effect and 
      mechanism of KLF4 in the carcinogenesis and progression of urothelial bladder 
      cancer (UBC). Immunohistochemistry (IHC) and quantitative RT-PCR (qRT-PCR) were 
      used to evaluate the expression of KLF4. The CpG methylation status of the 
      promoter region was analyzed using bisulfite-sequencing PCR (BSP). CRISPR-ON 
      system comprised sgRNA and dCas9 protein combined with a transcriptional 
      activation domain. The cell proliferation and cell cycle were assessed by CCK-8 
      assay, flow cytometry and colony formation assay. The cell motility ability was 
      evaluated using trans-well assay. In vivo tumorigenesis assay and lung metastasis 
      model were also performed. The KLF4 expression was significantly downregulated in 
      UBC tissues. The high CpG methylation status in the promoter of KLF4 was 
      confirmed using BSP. KLF4 overexpression was successfully achieved via CRISPR-ON 
      system, which inhibited the proliferation and induced G1-phase arrest in T24 
      cells through the regulation of AKT/p21 signal. Furthermore, enforced expression 
      of KLF4 also abrogated the migration and invasion of T24 cells by suppressing EMT 
      progression. Finally, in vivo models indicated that the upregulation of KLF4 
      could inhibit tumorigenesis and lung metastasis in nude mice. In conclusion, KLF4 
      overexpression mediated by CRISPR-ON inhibits tumorigenesis and EMT progression 
      in UBC cells, representing a potential therapeutic target, and CRISPR-ON system 
      could be a therapeutic strategy for UBC in the future.
FAU - Xu, Xin
AU  - Xu X
AD  - Department of Urology, The First Affiliated Hospital, School of Medicine, 
      Zhejiang University, Hangzhou 310003, Zhejiang Province, P.R. China.
FAU - Li, Jiangfeng
AU  - Li J
AD  - Department of Urology, The First Affiliated Hospital, School of Medicine, 
      Zhejiang University, Hangzhou 310003, Zhejiang Province, P.R. China.
FAU - Zhu, Yi
AU  - Zhu Y
AD  - Department of Urology, The First Affiliated Hospital, School of Medicine, 
      Zhejiang University, Hangzhou 310003, Zhejiang Province, P.R. China.
FAU - Xie, Bo
AU  - Xie B
AD  - Department of Urology, Tongde Hospital of Zhejiang Province, Hangzhou 310012, 
      Zhejiang Province, P.R. China.
FAU - Wang, Xiao
AU  - Wang X
AD  - Department of Urology, The First Affiliated Hospital, School of Medicine, 
      Zhejiang University, Hangzhou 310003, Zhejiang Province, P.R. China.
FAU - Wang, Song
AU  - Wang S
AD  - Department of Urology, The First Affiliated Hospital, School of Medicine, 
      Zhejiang University, Hangzhou 310003, Zhejiang Province, P.R. China.
FAU - Xie, Haiyun
AU  - Xie H
AD  - Department of Urology, The First Affiliated Hospital, School of Medicine, 
      Zhejiang University, Hangzhou 310003, Zhejiang Province, P.R. China.
FAU - Yan, Huaqing
AU  - Yan H
AD  - Department of Urology, The First Affiliated Hospital, School of Medicine, 
      Zhejiang University, Hangzhou 310003, Zhejiang Province, P.R. China.
FAU - Ying, Yufan
AU  - Ying Y
AD  - Department of Urology, The First Affiliated Hospital, School of Medicine, 
      Zhejiang University, Hangzhou 310003, Zhejiang Province, P.R. China.
FAU - Lin, Yiwei
AU  - Lin Y
AD  - Department of Urology, The First Affiliated Hospital, School of Medicine, 
      Zhejiang University, Hangzhou 310003, Zhejiang Province, P.R. China.
FAU - Liu, Ben
AU  - Liu B
AD  - Department of Urology, The First Affiliated Hospital, School of Medicine, 
      Zhejiang University, Hangzhou 310003, Zhejiang Province, P.R. China.
FAU - Wang, Wei
AU  - Wang W
AD  - Department of Urology, The First Affiliated Hospital, School of Medicine, 
      Zhejiang University, Hangzhou 310003, Zhejiang Province, P.R. China.
FAU - Zheng, Xiangyi
AU  - Zheng X
AD  - Department of Urology, The First Affiliated Hospital, School of Medicine, 
      Zhejiang University, Hangzhou 310003, Zhejiang Province, P.R. China.
LA  - eng
PT  - Journal Article
DEP - 20171027
PL  - United States
TA  - Oncotarget
JT  - Oncotarget
JID - 101532965
PMC - PMC5731936
OTO - NOTNLM
OT  - CRISPR-ON
OT  - EMT
OT  - KLF4
OT  - proliferation
OT  - urothelial bladder cancer
COIS- CONFLICTS OF INTEREST The authors declare no competing financial interests.
EDAT- 2017/12/20 06:00
MHDA- 2017/12/20 06:01
PMCR- 2017/11/24
CRDT- 2017/12/20 06:00
PHST- 2017/08/14 00:00 [received]
PHST- 2017/10/05 00:00 [accepted]
PHST- 2017/12/20 06:00 [entrez]
PHST- 2017/12/20 06:00 [pubmed]
PHST- 2017/12/20 06:01 [medline]
PHST- 2017/11/24 00:00 [pmc-release]
AID - 22158 [pii]
AID - 10.18632/oncotarget.22158 [doi]
PST - epublish
SO  - Oncotarget. 2017 Oct 27;8(60):102078-102087. doi: 10.18632/oncotarget.22158. 
      eCollection 2017 Nov 24.