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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...

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Autores principales: Tao, Yi-Jing, Li, Yong-ju, Zheng, Wen, Zhao, Juan-juan, Guo, Meng-meng, Zhou, Ya, Qin, Na-lin, Zheng, Jing, Xu, Lin
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
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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.
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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
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