Targeted Inhibitory Effect of Nasopharyngeal Carcinoma Cells by Hre 2 .Grp78 Chimeric Promoter Regulating Fusion Gene TK/VP3

• Published in: Technology in cancer research & treatment Vol. 18; p. 1533033819875166
• Main Authors: Li, Jin-Yun, Huang, Wen-Xiao, Chen, Jie, Zhao, Su-Ping, Tang, Yao-Yun
• Format: Journal Article
• Language: English
• Published: United States 01.01.2019
• Subjects: Apoptosis - genetics; Capsid Proteins - genetics; Capsid Proteins - metabolism; Cell Line, Tumor; Cell Proliferation - genetics; Gene Expression Regulation, Neoplastic; Heat-Shock Proteins - genetics; Humans; Hypoxia - genetics; Hypoxia - metabolism; Nasopharyngeal Carcinoma - genetics; Nasopharyngeal Carcinoma - metabolism; Plasmids - genetics; Promoter Regions, Genetic; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Response Elements; Thymidine Kinase - genetics; Thymidine Kinase - metabolism; Trans-Activators - genetics; Trans-Activators - metabolism; Viral Proteins - genetics; Viral Proteins - metabolism; VP3; plasmids construction; nasopharyngeal carcinoma; TK
• ISSN: 1533-0346; 1533-0338
• DOI: 10.1177/1533033819875166

To construct plasmids with Hre .Grp78 chimeric promoter regulating fusion gene and elaborate the effects of overexpressed on nasopharyngeal carcinoma cells. Four plasmids were constructed, including pcDNA3.1-CMV-TK/VP3, pcDNA3.1-Hre .TK/VP3, pcDNA3.1-Grp78.TK/VP3, and pcDNA3.1-Hre .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. 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 .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 .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. We successfully constructed 4 plasmids with Hre .Grp78 chimeric promoter regulating fusion gene which could significantly inhibit the proliferation as well as enhance the apoptosis of nasopharyngeal carcinoma cells under glucose deprivation or hypoxia condition.