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Targeted Inhibitory Effect of Nasopharyngeal Carcinoma Cells by Hre(2).Grp78 Chimeric Promoter Regulating Fusion Gene TK/VP3
OBJECTIVE: To construct plasmids with Hre(2).Grp78 chimeric promoter regulating fusion gene TK/VP3 and elaborate the effects of overexpressed TK/VP3 on nasopharyngeal carcinoma cells. METHODS: Four plasmids were constructed, including pcDNA3.1-CMV-TK/VP3, pcDNA3.1-Hre(2).TK/VP3, pcDNA3.1-Grp78.TK/VP...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
SAGE Publications
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6880038/ https://www.ncbi.nlm.nih.gov/pubmed/31769345 http://dx.doi.org/10.1177/1533033819875166 |
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author | Li, Jin-Yun Huang, Wen-Xiao Chen, Jie Zhao, Su-Ping Tang, Yao-Yun |
author_facet | Li, Jin-Yun Huang, Wen-Xiao Chen, Jie Zhao, Su-Ping Tang, Yao-Yun |
author_sort | Li, Jin-Yun |
collection | PubMed |
description | OBJECTIVE: To construct plasmids with Hre(2).Grp78 chimeric promoter regulating fusion gene TK/VP3 and elaborate the effects of overexpressed TK/VP3 on nasopharyngeal carcinoma cells. METHODS: Four plasmids were constructed, including pcDNA3.1-CMV-TK/VP3, pcDNA3.1-Hre(2).TK/VP3, pcDNA3.1-Grp78.TK/VP3, and pcDNA3.1-Hre(2).Grp78.TK/VP3. The human nasopharyngeal carcinoma cell line HNE1 cells were transfected with the 4 plasmids, respectively. Cell viabilities were evaluated using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and apoptosis was conducted using flow cytometry analysis. The expression of TK, VP3, Grp78, and hypoxia-inducible factor 1α and apoptosis-related proteins was determined by real-time quantitative polymerase chain reaction and Western blotting. RESULTS: The recombinant plasmids that could steadily overexpress TK and VP3 were successfully constructed. Expression of TK and VP3 in cells transfected with pcDNA3.1-Hre(2).TK/VP3 and pcDNA3.1-Grp78.TK/VP3 was significantly higher than pcDNA3.1-CMV-TK/VP3, and expression in cells transfected with pcDNA3.1-Hre(2).Grp78.TK/VP3 was the highest. Under glucose deprivation or hypoxia condition, Grp78 or hypoxia-inducible factor 1α was overexpressed so that expression of TK and VP3 was significantly upregulated, which could further inhibit cell proliferation and enhance cell apoptosis. CONCLUSION: We successfully constructed 4 plasmids with Hre(2).Grp78 chimeric promoter regulating fusion gene TK/VP3, which could significantly inhibit the proliferation as well as enhance the apoptosis of nasopharyngeal carcinoma cells under glucose deprivation or hypoxia condition. |
format | Online Article Text |
id | pubmed-6880038 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | SAGE Publications |
record_format | MEDLINE/PubMed |
spelling | pubmed-68800382019-12-05 Targeted Inhibitory Effect of Nasopharyngeal Carcinoma Cells by Hre(2).Grp78 Chimeric Promoter Regulating Fusion Gene TK/VP3 Li, Jin-Yun Huang, Wen-Xiao Chen, Jie Zhao, Su-Ping Tang, Yao-Yun Technol Cancer Res Treat Original Article OBJECTIVE: To construct plasmids with Hre(2).Grp78 chimeric promoter regulating fusion gene TK/VP3 and elaborate the effects of overexpressed TK/VP3 on nasopharyngeal carcinoma cells. METHODS: Four plasmids were constructed, including pcDNA3.1-CMV-TK/VP3, pcDNA3.1-Hre(2).TK/VP3, pcDNA3.1-Grp78.TK/VP3, and pcDNA3.1-Hre(2).Grp78.TK/VP3. The human nasopharyngeal carcinoma cell line HNE1 cells were transfected with the 4 plasmids, respectively. Cell viabilities were evaluated using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and apoptosis was conducted using flow cytometry analysis. The expression of TK, VP3, Grp78, and hypoxia-inducible factor 1α and apoptosis-related proteins was determined by real-time quantitative polymerase chain reaction and Western blotting. RESULTS: The recombinant plasmids that could steadily overexpress TK and VP3 were successfully constructed. Expression of TK and VP3 in cells transfected with pcDNA3.1-Hre(2).TK/VP3 and pcDNA3.1-Grp78.TK/VP3 was significantly higher than pcDNA3.1-CMV-TK/VP3, and expression in cells transfected with pcDNA3.1-Hre(2).Grp78.TK/VP3 was the highest. Under glucose deprivation or hypoxia condition, Grp78 or hypoxia-inducible factor 1α was overexpressed so that expression of TK and VP3 was significantly upregulated, which could further inhibit cell proliferation and enhance cell apoptosis. CONCLUSION: We successfully constructed 4 plasmids with Hre(2).Grp78 chimeric promoter regulating fusion gene TK/VP3, which could significantly inhibit the proliferation as well as enhance the apoptosis of nasopharyngeal carcinoma cells under glucose deprivation or hypoxia condition. SAGE Publications 2019-11-26 /pmc/articles/PMC6880038/ /pubmed/31769345 http://dx.doi.org/10.1177/1533033819875166 Text en © The Author(s) 2019 http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (http://www.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 Li, Jin-Yun Huang, Wen-Xiao Chen, Jie Zhao, Su-Ping Tang, Yao-Yun Targeted Inhibitory Effect of Nasopharyngeal Carcinoma Cells by Hre(2).Grp78 Chimeric Promoter Regulating Fusion Gene TK/VP3 |
title | Targeted Inhibitory Effect of Nasopharyngeal Carcinoma Cells by
Hre(2).Grp78 Chimeric Promoter Regulating Fusion Gene
TK/VP3 |
title_full | Targeted Inhibitory Effect of Nasopharyngeal Carcinoma Cells by
Hre(2).Grp78 Chimeric Promoter Regulating Fusion Gene
TK/VP3 |
title_fullStr | Targeted Inhibitory Effect of Nasopharyngeal Carcinoma Cells by
Hre(2).Grp78 Chimeric Promoter Regulating Fusion Gene
TK/VP3 |
title_full_unstemmed | Targeted Inhibitory Effect of Nasopharyngeal Carcinoma Cells by
Hre(2).Grp78 Chimeric Promoter Regulating Fusion Gene
TK/VP3 |
title_short | Targeted Inhibitory Effect of Nasopharyngeal Carcinoma Cells by
Hre(2).Grp78 Chimeric Promoter Regulating Fusion Gene
TK/VP3 |
title_sort | targeted inhibitory effect of nasopharyngeal carcinoma cells by
hre(2).grp78 chimeric promoter regulating fusion gene
tk/vp3 |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6880038/ https://www.ncbi.nlm.nih.gov/pubmed/31769345 http://dx.doi.org/10.1177/1533033819875166 |
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