title: MicroRNA-124-3p inhibits cell migration and invasion in bladder cancer cells by targeting ROCK1.
abstract: BACKGROUND
  Increasing evidence has suggested that dysregulation of certain microRNAs (miRNAs) may contribute to human disease including carcinogenesis and tumor metastasis in human. miR-124-3p is down-regulated in various cancers, and modulates proliferation and aggressiveness of cancer cells. However, the roles of miR-124-3p in human bladder cancer are elusive. Thus, this study was conducted to investigate the biological functions and its molecular mechanisms of miR-124-3p in human bladder cancer cell lines, discussing whether it has a potential to be a therapeutic biomarker of bladder cancer.
  
  
  METHODS
  Three human bladder cancer cell lines and samples from ten patients with bladder cancer were analyzed for the expression of miR-124-3p by quantitative RT--PCR. Exogenetic overexpression of miR-124-3p was established by transfecting mimics into T24, UM-UC-3 and J82 cells, after that cell proliferation and cell cycle were assessed by MTT assay, flow cytometry and Colony-forming assay. Cell motility and invasion ability were evaluated by wound healing assay and transwell assay. Tissue microarray, and immunohistochemistry with antibodies against ROCK1, MMP2 and MMP9 was performed using the peroxidase and DAB methods. The target gene of miR-124-3p was determined by luciferase assays, quantitative RT--PCR and western blot. The regulation of epithelial-to-mesenchymal transition by miR-124-3p was analyzed by western blot.
  
  
  RESULTS
  miR-124-3p is frequently down-regulated in bladder cancer both in three bladder cancer cell lines, T24, UM-UC-3, J82 and clinical samples. Overexpression of miR-124-3p induced G1-phase arrest in T24, UM-UC-3 and J82 cell lines and suppressed cell growth in colony-forming assay. miR-124-3p significantly repressed the capability of migration and invasion of bladder cancer cells. In addition, ROCK1 was identified as a new target of miR-124-3p. ROCK1, MMP2, MMP9 were up-regulated in bladder cancer tissues. Furthermore, we demonstrated miR-124-3p could inhibit bladder cancer cell epithelial mesenchymal transfer, and regulated the expression of c-Met, MMP2, MMP9.
  
  
  CONCLUSIONS
  miR-124-3p can repress the migration and invasion of bladder cancer cells via regulating ROCK1. Our data indicate that miR-124-3p could be a tumor suppressor and may have a potential to be a diagnostics or predictive biomarker in bladder cancer.
authors.forename: Xianglai
authors.initials: X
authors.lastname: Xu
authors.forename: Shiqi
authors.initials: S
authors.lastname: Li
authors.forename: Yiwei
authors.initials: Y
authors.lastname: Lin
authors.forename: Hong
authors.initials: H
authors.lastname: Chen
authors.forename: Zhenghui
authors.initials: Z
authors.lastname: Hu
authors.forename: Yeqing
authors.initials: Y
authors.lastname: Mao
authors.forename: Xin
authors.initials: X
authors.lastname: Xu
authors.forename: Jian
authors.initials: J
authors.lastname: Wu
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhu
authors.forename: Xiangyi
authors.initials: X
authors.lastname: Zheng
authors.forename: Jindan
authors.initials: J
authors.lastname: Luo
authors.affiliation: Department of Urology, the First Affiliated Hospital, Zhejiang University, Hangzhou, Zhejiang Province, China. xielp@zjuem.zju.edu.cn.
authors.forename: Liping
authors.initials: L
authors.lastname: Xie
mesh_terms: D020413:3' Untranslated Regions; D001483:Base Sequence; D015153:Blotting, Western; D002453:Cell Cycle; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology*; D017931:DNA Primers; D005434:Flow Cytometry; D006801:Humans; D035683:MicroRNAs / Q000502:physiology*; D009361:Neoplasm Invasiveness; D016133:Polymerase Chain Reaction; D001749:Urinary Bladder Neoplasms / Q000378:metabolism / Q000473:pathology*; D054460:rho-Associated Kinases / Q000235:genetics / Q000502:physiology*
chemical_list: D020413:3' Untranslated Regions; D017931:DNA Primers; C509207:MIRN124 microRNA, human; D035683:MicroRNAs; C515851:ROCK1 protein, human; D054460:rho-Associated Kinases