Cargando…
Antisense oligonucleotides against microRNA-21 reduced the proliferation and migration of human colon carcinoma cells
BACKGROUND: Colon carcinoma is one of the commonly tumors that threaten human beings as its highly morbidity and mortality. Recent evidences suggested that microRNA-21 (miR-21) played an important role in the development of colon carcinoma and might be a potential biological marker for the diagnosis...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2015
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4522075/ https://www.ncbi.nlm.nih.gov/pubmed/26236156 http://dx.doi.org/10.1186/s12935-015-0228-7 |
_version_ | 1782383908740923392 |
---|---|
author | Tao, Yi-Jing Li, Yong-ju Zheng, Wen Zhao, Juan-juan Guo, Meng-meng Zhou, Ya Qin, Na-lin Zheng, Jing Xu, Lin |
author_facet | Tao, Yi-Jing Li, Yong-ju Zheng, Wen Zhao, Juan-juan Guo, Meng-meng Zhou, Ya Qin, Na-lin Zheng, Jing Xu, Lin |
author_sort | Tao, Yi-Jing |
collection | PubMed |
description | BACKGROUND: Colon carcinoma is one of the commonly tumors that threaten human beings as its highly morbidity and mortality. Recent evidences suggested that microRNA-21 (miR-21) played an important role in the development of colon carcinoma and might be a potential biological marker for the diagnosis and prognosis of colon carcinoma. However, the potential effect of miR-21 based therapeutic studies in colon carcinoma remains to be fully elucidated. METHODS: In present study, we constructed an eukaryotic expression vector encoding antisense oligonucleotides against miR-21 (termed as p-miR-21-ASO) and the expression of miRNA-21 in human colon cancer was detected by Real-time PCR. To assess its possible effect on the proliferation and migration capacity of human colon carcinoma cells in vitro, CCK-8 assay, colony formation assay and cell invasion, as well as migration assay, were performed respectively. Moreover, PTEN, one of target molecules of miRNA-21, was analyzed by Western blot and Fluorescence activated cell sorter assay. Finally, the transduction of AKT and ERK pathways in human colon carcinoma cells was determined by Western blot. RESULTS: We found that transiently transfection of p-miR-21-ASO could efficiently decrease the relative expression of miR-21 in human colon carcinoma HCT116 cells, accompanied by impaired proliferation and clone formation. Furthermore, we found that down-regulation of miR-21 also could significantly abrogate the invasion and migration capacity in vitro, as well as the expression of vascular endothelial growth factor which is critical for the metastatic capacity of colon carcinoma cells. Mechanistic evidence showed that down-regulation of miR-21 increased the expression of its target molecule PTEN in HCT116 cells. Finally, we revealed that the expression level of both phosphor-ERK1/2 and phosphor-AKT also were altered. CONCLUSIONS: Therefore, our data suggested miR-21 ASO against miR-21 might be a useful strategy to alter the expression of miR-21 in colon carcinoma cells, which was helpful for the development of miR-21-based therapeutic strategies against clinical colon carcinoma. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12935-015-0228-7) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-4522075 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-45220752015-08-02 Antisense oligonucleotides against microRNA-21 reduced the proliferation and migration of human colon carcinoma cells Tao, Yi-Jing Li, Yong-ju Zheng, Wen Zhao, Juan-juan Guo, Meng-meng Zhou, Ya Qin, Na-lin Zheng, Jing Xu, Lin Cancer Cell Int Primary Research BACKGROUND: Colon carcinoma is one of the commonly tumors that threaten human beings as its highly morbidity and mortality. Recent evidences suggested that microRNA-21 (miR-21) played an important role in the development of colon carcinoma and might be a potential biological marker for the diagnosis and prognosis of colon carcinoma. However, the potential effect of miR-21 based therapeutic studies in colon carcinoma remains to be fully elucidated. METHODS: In present study, we constructed an eukaryotic expression vector encoding antisense oligonucleotides against miR-21 (termed as p-miR-21-ASO) and the expression of miRNA-21 in human colon cancer was detected by Real-time PCR. To assess its possible effect on the proliferation and migration capacity of human colon carcinoma cells in vitro, CCK-8 assay, colony formation assay and cell invasion, as well as migration assay, were performed respectively. Moreover, PTEN, one of target molecules of miRNA-21, was analyzed by Western blot and Fluorescence activated cell sorter assay. Finally, the transduction of AKT and ERK pathways in human colon carcinoma cells was determined by Western blot. RESULTS: We found that transiently transfection of p-miR-21-ASO could efficiently decrease the relative expression of miR-21 in human colon carcinoma HCT116 cells, accompanied by impaired proliferation and clone formation. Furthermore, we found that down-regulation of miR-21 also could significantly abrogate the invasion and migration capacity in vitro, as well as the expression of vascular endothelial growth factor which is critical for the metastatic capacity of colon carcinoma cells. Mechanistic evidence showed that down-regulation of miR-21 increased the expression of its target molecule PTEN in HCT116 cells. Finally, we revealed that the expression level of both phosphor-ERK1/2 and phosphor-AKT also were altered. CONCLUSIONS: Therefore, our data suggested miR-21 ASO against miR-21 might be a useful strategy to alter the expression of miR-21 in colon carcinoma cells, which was helpful for the development of miR-21-based therapeutic strategies against clinical colon carcinoma. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12935-015-0228-7) contains supplementary material, which is available to authorized users. BioMed Central 2015-08-01 /pmc/articles/PMC4522075/ /pubmed/26236156 http://dx.doi.org/10.1186/s12935-015-0228-7 Text en © Tao et al. 2015 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Primary Research Tao, Yi-Jing Li, Yong-ju Zheng, Wen Zhao, Juan-juan Guo, Meng-meng Zhou, Ya Qin, Na-lin Zheng, Jing Xu, Lin Antisense oligonucleotides against microRNA-21 reduced the proliferation and migration of human colon carcinoma cells |
title | Antisense oligonucleotides against microRNA-21 reduced the proliferation and migration of human colon carcinoma cells |
title_full | Antisense oligonucleotides against microRNA-21 reduced the proliferation and migration of human colon carcinoma cells |
title_fullStr | Antisense oligonucleotides against microRNA-21 reduced the proliferation and migration of human colon carcinoma cells |
title_full_unstemmed | Antisense oligonucleotides against microRNA-21 reduced the proliferation and migration of human colon carcinoma cells |
title_short | Antisense oligonucleotides against microRNA-21 reduced the proliferation and migration of human colon carcinoma cells |
title_sort | antisense oligonucleotides against microrna-21 reduced the proliferation and migration of human colon carcinoma cells |
topic | Primary Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4522075/ https://www.ncbi.nlm.nih.gov/pubmed/26236156 http://dx.doi.org/10.1186/s12935-015-0228-7 |
work_keys_str_mv | AT taoyijing antisenseoligonucleotidesagainstmicrorna21reducedtheproliferationandmigrationofhumancoloncarcinomacells AT liyongju antisenseoligonucleotidesagainstmicrorna21reducedtheproliferationandmigrationofhumancoloncarcinomacells AT zhengwen antisenseoligonucleotidesagainstmicrorna21reducedtheproliferationandmigrationofhumancoloncarcinomacells AT zhaojuanjuan antisenseoligonucleotidesagainstmicrorna21reducedtheproliferationandmigrationofhumancoloncarcinomacells AT guomengmeng antisenseoligonucleotidesagainstmicrorna21reducedtheproliferationandmigrationofhumancoloncarcinomacells AT zhouya antisenseoligonucleotidesagainstmicrorna21reducedtheproliferationandmigrationofhumancoloncarcinomacells AT qinnalin antisenseoligonucleotidesagainstmicrorna21reducedtheproliferationandmigrationofhumancoloncarcinomacells AT zhengjing antisenseoligonucleotidesagainstmicrorna21reducedtheproliferationandmigrationofhumancoloncarcinomacells AT xulin antisenseoligonucleotidesagainstmicrorna21reducedtheproliferationandmigrationofhumancoloncarcinomacells |