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Restoration of miR-20a expression suppresses cell proliferation, migration, and invasion in HepG2 cells
OBJECTIVE: To study microRNA (miR)-20a expression in hepatocellular carcinoma (HCC) and its effects on the proliferation, migration, and invasion of HepG2. METHODS: The real-time polymerase chain reaction was used to detect the expression of miR-20a in HCC tissue and normal tissue, as well as in HCC...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Dove Medical Press
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4892835/ https://www.ncbi.nlm.nih.gov/pubmed/27313460 http://dx.doi.org/10.2147/OTT.S96861 |
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author | Chen, Guang Shun Zhou, Ning Li, Jie-Qun Li, Ting Zhang, Zhong-Qiang Si, Zhong-Zhou |
author_facet | Chen, Guang Shun Zhou, Ning Li, Jie-Qun Li, Ting Zhang, Zhong-Qiang Si, Zhong-Zhou |
author_sort | Chen, Guang Shun |
collection | PubMed |
description | OBJECTIVE: To study microRNA (miR)-20a expression in hepatocellular carcinoma (HCC) and its effects on the proliferation, migration, and invasion of HepG2. METHODS: The real-time polymerase chain reaction was used to detect the expression of miR-20a in HCC tissue and normal tissue, as well as in HCC cell lines and normal liver cells. miR-20a mimic and miR negative control (NC) were transfected into HepG2 cells. MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) assay was used to detect cell proliferation. Annexin fluorescein isothiocyanate/propidium iodide assay was run to examine the early apoptosis of cells. Transwell chamber assay was carried out to investigate the cell invasion and migration abilities. RESULTS: miR-20a was lowly expressed both in HCC tissues and HCC cell lines. After transfection of exogenous miR-20 mimics, miR-20a expression in HepG2 cells was significantly increased by 61.29% compared to the blank group (P<0.01). MTT assay showed that the growth of HepG2 cells in the miR-20a mimics group was significantly inhibited, and optical density values during the 36–96 hour time period were dramatically decreased compared to the blank group (P<0.01). Apoptosis rates of the miR-20a mimics group were higher than those of the blank and NC groups (both P<0.01). The number of HCC cells after transfection by miR-20a mimics in the G1 and S phases were 15.88% and 7.89%, respectively, which were lower than in the blank and NC groups (both P<0.05). Transwell assay showed that in the miR-20a mimics group the number of cell migration and invasion were 0.459 and 0.501 times that of the blank group (both P<0.01), and the migration and inhibition rates were 54.1% and 51.4%, respectively. After closing target gene CCND1 in HepG2 cells, the number of cell migration and invasion in the small interfering (si)-CCND1 group were 0.444 and 0.435 times that of the si-NC group (P<0.05); and compared to the si-NC group, the migration and inhibition rates were 55.6% and 56.5%, respectively. CONCLUSION: miR-20a can inhibit the growth, invasion, and migration of HepG2 cells, and is therefore promising as a new molecular target for diagnosis and therapy of HCC. |
format | Online Article Text |
id | pubmed-4892835 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Dove Medical Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-48928352016-06-16 Restoration of miR-20a expression suppresses cell proliferation, migration, and invasion in HepG2 cells Chen, Guang Shun Zhou, Ning Li, Jie-Qun Li, Ting Zhang, Zhong-Qiang Si, Zhong-Zhou Onco Targets Ther Original Research OBJECTIVE: To study microRNA (miR)-20a expression in hepatocellular carcinoma (HCC) and its effects on the proliferation, migration, and invasion of HepG2. METHODS: The real-time polymerase chain reaction was used to detect the expression of miR-20a in HCC tissue and normal tissue, as well as in HCC cell lines and normal liver cells. miR-20a mimic and miR negative control (NC) were transfected into HepG2 cells. MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) assay was used to detect cell proliferation. Annexin fluorescein isothiocyanate/propidium iodide assay was run to examine the early apoptosis of cells. Transwell chamber assay was carried out to investigate the cell invasion and migration abilities. RESULTS: miR-20a was lowly expressed both in HCC tissues and HCC cell lines. After transfection of exogenous miR-20 mimics, miR-20a expression in HepG2 cells was significantly increased by 61.29% compared to the blank group (P<0.01). MTT assay showed that the growth of HepG2 cells in the miR-20a mimics group was significantly inhibited, and optical density values during the 36–96 hour time period were dramatically decreased compared to the blank group (P<0.01). Apoptosis rates of the miR-20a mimics group were higher than those of the blank and NC groups (both P<0.01). The number of HCC cells after transfection by miR-20a mimics in the G1 and S phases were 15.88% and 7.89%, respectively, which were lower than in the blank and NC groups (both P<0.05). Transwell assay showed that in the miR-20a mimics group the number of cell migration and invasion were 0.459 and 0.501 times that of the blank group (both P<0.01), and the migration and inhibition rates were 54.1% and 51.4%, respectively. After closing target gene CCND1 in HepG2 cells, the number of cell migration and invasion in the small interfering (si)-CCND1 group were 0.444 and 0.435 times that of the si-NC group (P<0.05); and compared to the si-NC group, the migration and inhibition rates were 55.6% and 56.5%, respectively. CONCLUSION: miR-20a can inhibit the growth, invasion, and migration of HepG2 cells, and is therefore promising as a new molecular target for diagnosis and therapy of HCC. Dove Medical Press 2016-05-27 /pmc/articles/PMC4892835/ /pubmed/27313460 http://dx.doi.org/10.2147/OTT.S96861 Text en © 2016 Chen et al. This work is published and licensed by Dove Medical Press Limited The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. |
spellingShingle | Original Research Chen, Guang Shun Zhou, Ning Li, Jie-Qun Li, Ting Zhang, Zhong-Qiang Si, Zhong-Zhou Restoration of miR-20a expression suppresses cell proliferation, migration, and invasion in HepG2 cells |
title | Restoration of miR-20a expression suppresses cell proliferation, migration, and invasion in HepG2 cells |
title_full | Restoration of miR-20a expression suppresses cell proliferation, migration, and invasion in HepG2 cells |
title_fullStr | Restoration of miR-20a expression suppresses cell proliferation, migration, and invasion in HepG2 cells |
title_full_unstemmed | Restoration of miR-20a expression suppresses cell proliferation, migration, and invasion in HepG2 cells |
title_short | Restoration of miR-20a expression suppresses cell proliferation, migration, and invasion in HepG2 cells |
title_sort | restoration of mir-20a expression suppresses cell proliferation, migration, and invasion in hepg2 cells |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4892835/ https://www.ncbi.nlm.nih.gov/pubmed/27313460 http://dx.doi.org/10.2147/OTT.S96861 |
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