Gantao Chen,Yanxiang Cheng,Xiao Yang,Liu Yaodan,Weiguo Dong. Targeting of RhoE inhibits epithelial-mesenchymal transition during colorectal cancer cell migration. Oncol Transl Med, 2016, 2: 119-126.
Targeting of RhoE inhibits epithelial-mesenchymal transition during colorectal cancer cell migration
Received:December 14, 2015  Revised:June 10, 2016
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KeyWord:miR-200b; colorectal cancer (CRC); metalloproteinase (MMP); epithelial-mesenchymal transition (EMT); cell migration
Author NameAffiliationE-mail
Gantao Chen Renmin Hospital of Wuhan University 1226076952@qq.com 
Yanxiang Cheng Renmin Hospital of Wuhan University doctornancy@qq.com 
Xiao Yang Renmin Hospital of Wuhan University yangxiao_5200@163.com 
Liu Yaodan Renmin hospital of Wuhan University 1228965422@qq.com 
Weiguo Dong Renmin Hospital of Wuhan University 415100331@qq.com 
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Abstract:
      Objective: Despite microRNA (miR-200b) being proved to promote the proliferation of colorectal cancer (CRC) cells, the relationship between miR-200b and epithelial-mesenchymal transition (EMT) of CRC cells remains poorly understood. The aim of the study was to investigate the relationship between miR-200b and EMT during CRC cell migration. Methods: The effect of miR-200b on EMT-associated markers E-cadherin and vimentin was evaluated by western blot in CRC cells (SW620 and HT-29) by treatment with miR-200b mimics and inhibitors. A luciferase reporter assay was employed to detect downstream targets of miR-200b. Transwell migration assays were used to detect CRC cell migration. Results: Western blots revealed that treatment with miR-200b mimics led to up-regulation of E-cadherin and down-regulation of vimentin, metalloproteinase (MMP)-9, and MMP-2, whereas treatment with miR- 200b inhibitor exhibited opposite effects on expression of E-cadherin and vimentin. Luciferase reporter assays demonstrated that RhoE (RND3) was targeted by miR-200b. Two predicted target sites of miR-200b were present in the 3’-UTR of RhoE. Predicted target site 1 was from nucleotides 1584 to 1591, and site 2 was from nucleotides 1729 to 1735. RhoE knockdown cell lines were also established to investigate the impact of RhoE and miR-200b on EMT and cell migration. RhoE knockdown enhanced the effect of miR- 200b mimics, up-regulating E-cadherin and down-regulating vimentin. RhoE knockdown also inhibited cell migration. Furthermore, miR-200b mimic treatment further promoted the inhibitory effect of RhoE knockdown on cell migration. Conclusion: miR-200b inhibited EMT and CRC cell migration partly via inhibiting RhoE expression in CRC. RhoE and miR-200b might therefore be promising target genes in the management of CRC.
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