Cargando…

MicroRNA-124-3p inhibits cell migration and invasion in bladder cancer cells by targeting ROCK1

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. Howe...

Descripción completa

Detalles Bibliográficos
Autores principales: Xu, Xianglai, Li, Shiqi, Lin, Yiwei, Chen, Hong, Hu, Zhenghui, Mao, Yeqing, Xu, Xin, Wu, Jian, Zhu, Yi, Zheng, Xiangyi, Luo, Jindan, Xie, Liping
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4228407/
https://www.ncbi.nlm.nih.gov/pubmed/24180482
http://dx.doi.org/10.1186/1479-5876-11-276
_version_ 1782343976828796928
author Xu, Xianglai
Li, Shiqi
Lin, Yiwei
Chen, Hong
Hu, Zhenghui
Mao, Yeqing
Xu, Xin
Wu, Jian
Zhu, Yi
Zheng, Xiangyi
Luo, Jindan
Xie, Liping
author_facet Xu, Xianglai
Li, Shiqi
Lin, Yiwei
Chen, Hong
Hu, Zhenghui
Mao, Yeqing
Xu, Xin
Wu, Jian
Zhu, Yi
Zheng, Xiangyi
Luo, Jindan
Xie, Liping
author_sort Xu, Xianglai
collection PubMed
description 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.
format Online
Article
Text
id pubmed-4228407
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-42284072014-11-13 MicroRNA-124-3p inhibits cell migration and invasion in bladder cancer cells by targeting ROCK1 Xu, Xianglai Li, Shiqi Lin, Yiwei Chen, Hong Hu, Zhenghui Mao, Yeqing Xu, Xin Wu, Jian Zhu, Yi Zheng, Xiangyi Luo, Jindan Xie, Liping J Transl Med Research 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. BioMed Central 2013-11-02 /pmc/articles/PMC4228407/ /pubmed/24180482 http://dx.doi.org/10.1186/1479-5876-11-276 Text en Copyright © 2013 Xu et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Xu, Xianglai
Li, Shiqi
Lin, Yiwei
Chen, Hong
Hu, Zhenghui
Mao, Yeqing
Xu, Xin
Wu, Jian
Zhu, Yi
Zheng, Xiangyi
Luo, Jindan
Xie, Liping
MicroRNA-124-3p inhibits cell migration and invasion in bladder cancer cells by targeting ROCK1
title MicroRNA-124-3p inhibits cell migration and invasion in bladder cancer cells by targeting ROCK1
title_full MicroRNA-124-3p inhibits cell migration and invasion in bladder cancer cells by targeting ROCK1
title_fullStr MicroRNA-124-3p inhibits cell migration and invasion in bladder cancer cells by targeting ROCK1
title_full_unstemmed MicroRNA-124-3p inhibits cell migration and invasion in bladder cancer cells by targeting ROCK1
title_short MicroRNA-124-3p inhibits cell migration and invasion in bladder cancer cells by targeting ROCK1
title_sort microrna-124-3p inhibits cell migration and invasion in bladder cancer cells by targeting rock1
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4228407/
https://www.ncbi.nlm.nih.gov/pubmed/24180482
http://dx.doi.org/10.1186/1479-5876-11-276
work_keys_str_mv AT xuxianglai microrna1243pinhibitscellmigrationandinvasioninbladdercancercellsbytargetingrock1
AT lishiqi microrna1243pinhibitscellmigrationandinvasioninbladdercancercellsbytargetingrock1
AT linyiwei microrna1243pinhibitscellmigrationandinvasioninbladdercancercellsbytargetingrock1
AT chenhong microrna1243pinhibitscellmigrationandinvasioninbladdercancercellsbytargetingrock1
AT huzhenghui microrna1243pinhibitscellmigrationandinvasioninbladdercancercellsbytargetingrock1
AT maoyeqing microrna1243pinhibitscellmigrationandinvasioninbladdercancercellsbytargetingrock1
AT xuxin microrna1243pinhibitscellmigrationandinvasioninbladdercancercellsbytargetingrock1
AT wujian microrna1243pinhibitscellmigrationandinvasioninbladdercancercellsbytargetingrock1
AT zhuyi microrna1243pinhibitscellmigrationandinvasioninbladdercancercellsbytargetingrock1
AT zhengxiangyi microrna1243pinhibitscellmigrationandinvasioninbladdercancercellsbytargetingrock1
AT luojindan microrna1243pinhibitscellmigrationandinvasioninbladdercancercellsbytargetingrock1
AT xieliping microrna1243pinhibitscellmigrationandinvasioninbladdercancercellsbytargetingrock1