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MicroRNA-126-5p Inhibits the Migration of Breast Cancer Cells by Directly Targeting CNOT7
BACKGROUND: To assess the effect of microRNA-126-5p (miR-126-5p) on the migration of the breast cancer MCF7 cell line. METHODS: GSE143564 was downloaded from the Gene Expression Omnibus (GEO; http://www.ncbi.nlm.nih.gov/geo) to identify the differentially expressed miRNAs between breast cancer and a...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
SAGE Publications
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7711228/ https://www.ncbi.nlm.nih.gov/pubmed/33256566 http://dx.doi.org/10.1177/1533033820977545 |
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author | Miao, Yuying Lu, Jiang Fan, Baozhen Sun, Lecan |
author_facet | Miao, Yuying Lu, Jiang Fan, Baozhen Sun, Lecan |
author_sort | Miao, Yuying |
collection | PubMed |
description | BACKGROUND: To assess the effect of microRNA-126-5p (miR-126-5p) on the migration of the breast cancer MCF7 cell line. METHODS: GSE143564 was downloaded from the Gene Expression Omnibus (GEO; http://www.ncbi.nlm.nih.gov/geo) to identify the differentially expressed miRNAs between breast cancer and adjacent tissues. Quantitative reverse transcription PCR (RT-qPCR) was used to assess miR-126-5p levels in the normal 184A1 breast cell line and the breast cancer MCF7 cell line. The MCF7 cell line was then transfected with miR-126-5p mimics or corresponding negative control (NC-mimic). The proliferation and migration abilities of the MCF7 cell line were measured by methyl thiazolyl tetrazolium (MTT), Transwell and scratch healing assays. CCR4-NOT transcription complex and subunit 7 (CNOT7) expression levels in the NC-mimic and miR-126-5p mimic groups were measured by Western blot analysis. Bioinformatic analysis and a dual-luciferase reporter assay were performed to identify the miR-126-5p target gene. RESULTS: One hundred forty-eight differentially expressed miRNAs (downregulated = 55, upregulated = 93) were identified. MiR-126-5p expression in the MCF7 cell line was significantly downregulated relative to that of 184A1 cell line (P < 0.05). Compared with that observed in the control and NC-mimic groups, cell proliferation in the miR-126-5p mimic group was significantly decreased at 48 and 72 h posttransfection (P < 0.05). In addition, the scratch healing rate and number of membrane-piercing cells in the miR-126-5p overexpression group were lower than those detected in the control and NC groups (P < 0.05). Furthermore, miR-126-5p could reduce the luciferase activity for the wild-type CNOT7 gene 3’-untranslated region (UTR) reporter (P < 0.05) but had no effect on the mutant 3’UTR reporter (P > 0.05). Compared with that observed in the NC and control groups, the levels of CNOT7 in the miR-126-5p overexpression group decreased (P < 0.05). CONCLUSION: Upregulation of miR-126-5p can inhibit the migration of the breast cancer MCF7 cell line, which may involve its direct targeting of the 3’UTR of CNOT7. |
format | Online Article Text |
id | pubmed-7711228 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | SAGE Publications |
record_format | MEDLINE/PubMed |
spelling | pubmed-77112282020-12-08 MicroRNA-126-5p Inhibits the Migration of Breast Cancer Cells by Directly Targeting CNOT7 Miao, Yuying Lu, Jiang Fan, Baozhen Sun, Lecan Technol Cancer Res Treat Original Article BACKGROUND: To assess the effect of microRNA-126-5p (miR-126-5p) on the migration of the breast cancer MCF7 cell line. METHODS: GSE143564 was downloaded from the Gene Expression Omnibus (GEO; http://www.ncbi.nlm.nih.gov/geo) to identify the differentially expressed miRNAs between breast cancer and adjacent tissues. Quantitative reverse transcription PCR (RT-qPCR) was used to assess miR-126-5p levels in the normal 184A1 breast cell line and the breast cancer MCF7 cell line. The MCF7 cell line was then transfected with miR-126-5p mimics or corresponding negative control (NC-mimic). The proliferation and migration abilities of the MCF7 cell line were measured by methyl thiazolyl tetrazolium (MTT), Transwell and scratch healing assays. CCR4-NOT transcription complex and subunit 7 (CNOT7) expression levels in the NC-mimic and miR-126-5p mimic groups were measured by Western blot analysis. Bioinformatic analysis and a dual-luciferase reporter assay were performed to identify the miR-126-5p target gene. RESULTS: One hundred forty-eight differentially expressed miRNAs (downregulated = 55, upregulated = 93) were identified. MiR-126-5p expression in the MCF7 cell line was significantly downregulated relative to that of 184A1 cell line (P < 0.05). Compared with that observed in the control and NC-mimic groups, cell proliferation in the miR-126-5p mimic group was significantly decreased at 48 and 72 h posttransfection (P < 0.05). In addition, the scratch healing rate and number of membrane-piercing cells in the miR-126-5p overexpression group were lower than those detected in the control and NC groups (P < 0.05). Furthermore, miR-126-5p could reduce the luciferase activity for the wild-type CNOT7 gene 3’-untranslated region (UTR) reporter (P < 0.05) but had no effect on the mutant 3’UTR reporter (P > 0.05). Compared with that observed in the NC and control groups, the levels of CNOT7 in the miR-126-5p overexpression group decreased (P < 0.05). CONCLUSION: Upregulation of miR-126-5p can inhibit the migration of the breast cancer MCF7 cell line, which may involve its direct targeting of the 3’UTR of CNOT7. SAGE Publications 2020-12-01 /pmc/articles/PMC7711228/ /pubmed/33256566 http://dx.doi.org/10.1177/1533033820977545 Text en © The Author(s) 2020 https://creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage). |
spellingShingle | Original Article Miao, Yuying Lu, Jiang Fan, Baozhen Sun, Lecan MicroRNA-126-5p Inhibits the Migration of Breast Cancer Cells by Directly Targeting CNOT7 |
title | MicroRNA-126-5p Inhibits the Migration of Breast Cancer Cells by
Directly Targeting CNOT7 |
title_full | MicroRNA-126-5p Inhibits the Migration of Breast Cancer Cells by
Directly Targeting CNOT7 |
title_fullStr | MicroRNA-126-5p Inhibits the Migration of Breast Cancer Cells by
Directly Targeting CNOT7 |
title_full_unstemmed | MicroRNA-126-5p Inhibits the Migration of Breast Cancer Cells by
Directly Targeting CNOT7 |
title_short | MicroRNA-126-5p Inhibits the Migration of Breast Cancer Cells by
Directly Targeting CNOT7 |
title_sort | microrna-126-5p inhibits the migration of breast cancer cells by
directly targeting cnot7 |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7711228/ https://www.ncbi.nlm.nih.gov/pubmed/33256566 http://dx.doi.org/10.1177/1533033820977545 |
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