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Functions of MiRNA-128 on the Regulation of Head and Neck Squamous Cell Carcinoma Growth and Apoptosis
BACKGROUND: Incidence of head and neck squamous cell carcinoma (HNSCC) has continuously increased in past years while its survival rate has not been significantly improved. There is a critical need to better understand the genetic regulation of HNSCC tumorigenesis and progression. In this study, we...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4357443/ https://www.ncbi.nlm.nih.gov/pubmed/25764126 http://dx.doi.org/10.1371/journal.pone.0116321 |
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author | Hauser, Belinda Zhao, Yuan Pang, Xiaowu Ling, Zhiqiang Myers, Ernest Wang, Paul Califano, Joseph Gu, Xinbin |
author_facet | Hauser, Belinda Zhao, Yuan Pang, Xiaowu Ling, Zhiqiang Myers, Ernest Wang, Paul Califano, Joseph Gu, Xinbin |
author_sort | Hauser, Belinda |
collection | PubMed |
description | BACKGROUND: Incidence of head and neck squamous cell carcinoma (HNSCC) has continuously increased in past years while its survival rate has not been significantly improved. There is a critical need to better understand the genetic regulation of HNSCC tumorigenesis and progression. In this study, we comprehensively analyzed the function of miRNA-128 (miR-128) in the regulation of HNSCC growth and its putative targets in vitro and in vivo systems. METHODS: The function and targets of miR-128 were investigated in human HNSCC cell lines (JHU-13 and JHU-22), which were stably transfected with the miR-128 gene using a lentiviral delivery system. The expression levels of miR-128 and its targeted proteins were analyzed with qRT-PCR, Western blotting and flow cytometry. The binding capacity of miRNA-128 to its putative targets was determined using a luciferase report assay. MTT, colony formation, and a tumor xenograft model further evaluated the effects of miR-128 on HNSCC growth. RESULTS: We generated two miR-128 stably transfected human HNSCC cell lines (JHU-13(miR-128) and JHU-22(miR-128)). Enforced expression of miR-128 was detected in both cultured JHU-13(miR-128) and JHU-22(miR-128) cell lines, approximately seventeen to twenty folds higher than in vector control cell lines. miRNA-128 was able to bind with the 3′-untranslated regions of BMI-1, BAG-2, BAX, H3f3b, and Paip2 mRNAs, resulting in significant reduction of the targeted protein levels. We found that upregulated miR-128 expression significantly inhibited both JHU-13(miR-128) and JHU-22(miR-128) cell viability approximately 20 to 40%, and the JHU-22(miR-128) tumor xenograft growth compared to the vector control groups. CONCLUSIONS: miR-128 acted as a tumor suppressor inhibiting the HNSCC growth by directly mediating the expression of putative targets. Our results provide a better understanding of miRNA-128 function and its potential targets, which may be valuable for developing novel diagnostic markers and targeted therapy. |
format | Online Article Text |
id | pubmed-4357443 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-43574432015-03-23 Functions of MiRNA-128 on the Regulation of Head and Neck Squamous Cell Carcinoma Growth and Apoptosis Hauser, Belinda Zhao, Yuan Pang, Xiaowu Ling, Zhiqiang Myers, Ernest Wang, Paul Califano, Joseph Gu, Xinbin PLoS One Research Article BACKGROUND: Incidence of head and neck squamous cell carcinoma (HNSCC) has continuously increased in past years while its survival rate has not been significantly improved. There is a critical need to better understand the genetic regulation of HNSCC tumorigenesis and progression. In this study, we comprehensively analyzed the function of miRNA-128 (miR-128) in the regulation of HNSCC growth and its putative targets in vitro and in vivo systems. METHODS: The function and targets of miR-128 were investigated in human HNSCC cell lines (JHU-13 and JHU-22), which were stably transfected with the miR-128 gene using a lentiviral delivery system. The expression levels of miR-128 and its targeted proteins were analyzed with qRT-PCR, Western blotting and flow cytometry. The binding capacity of miRNA-128 to its putative targets was determined using a luciferase report assay. MTT, colony formation, and a tumor xenograft model further evaluated the effects of miR-128 on HNSCC growth. RESULTS: We generated two miR-128 stably transfected human HNSCC cell lines (JHU-13(miR-128) and JHU-22(miR-128)). Enforced expression of miR-128 was detected in both cultured JHU-13(miR-128) and JHU-22(miR-128) cell lines, approximately seventeen to twenty folds higher than in vector control cell lines. miRNA-128 was able to bind with the 3′-untranslated regions of BMI-1, BAG-2, BAX, H3f3b, and Paip2 mRNAs, resulting in significant reduction of the targeted protein levels. We found that upregulated miR-128 expression significantly inhibited both JHU-13(miR-128) and JHU-22(miR-128) cell viability approximately 20 to 40%, and the JHU-22(miR-128) tumor xenograft growth compared to the vector control groups. CONCLUSIONS: miR-128 acted as a tumor suppressor inhibiting the HNSCC growth by directly mediating the expression of putative targets. Our results provide a better understanding of miRNA-128 function and its potential targets, which may be valuable for developing novel diagnostic markers and targeted therapy. Public Library of Science 2015-03-12 /pmc/articles/PMC4357443/ /pubmed/25764126 http://dx.doi.org/10.1371/journal.pone.0116321 Text en © 2015 Hauser et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Hauser, Belinda Zhao, Yuan Pang, Xiaowu Ling, Zhiqiang Myers, Ernest Wang, Paul Califano, Joseph Gu, Xinbin Functions of MiRNA-128 on the Regulation of Head and Neck Squamous Cell Carcinoma Growth and Apoptosis |
title | Functions of MiRNA-128 on the Regulation of Head and Neck Squamous Cell Carcinoma Growth and Apoptosis |
title_full | Functions of MiRNA-128 on the Regulation of Head and Neck Squamous Cell Carcinoma Growth and Apoptosis |
title_fullStr | Functions of MiRNA-128 on the Regulation of Head and Neck Squamous Cell Carcinoma Growth and Apoptosis |
title_full_unstemmed | Functions of MiRNA-128 on the Regulation of Head and Neck Squamous Cell Carcinoma Growth and Apoptosis |
title_short | Functions of MiRNA-128 on the Regulation of Head and Neck Squamous Cell Carcinoma Growth and Apoptosis |
title_sort | functions of mirna-128 on the regulation of head and neck squamous cell carcinoma growth and apoptosis |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4357443/ https://www.ncbi.nlm.nih.gov/pubmed/25764126 http://dx.doi.org/10.1371/journal.pone.0116321 |
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