[Killing effect of VP3 on human nasopharyngeal carcinoma cell line CNE-2 cells].
Jing Xu, Yuanzheng Qiu, Yao‐Yun Tang, Yongquan Tian, Xianzhong Xiao, Suping Zhao
Abstract
Jing Xu, Yuanzheng Qiu, Yao‐Yun Tang, Yongquan Tian, Xianzhong Xiao, Suping Zhao
Abstract
OBJECTIVE: To investigate the killing effects of VP(3) on nasopharyngeal carcinoma cell line CNE-2. METHODS: Plasmid expression vector pcDNA3.1(-) CMV.VP(3)-His was constructed and identified by Kpn I/EcoR I endonuclease analysis, and then sequenced to verify successful insertion in the sense direction of VP(3) gene. pcDNA3.1(-) CMV.VP(3)-His and pcDNA3.1(-)-His expression plasmid was transiently transfected into nasopharyngeal carcinoma cell line CNE-2 . VP(3) protein expression was detected by Western blotting. MTT assay was used to determine the killing effects of VP(3) gene on nasopharyngeal carcinoma cell line CNE-2. RESULTS: Endonuclease analysis and sequencing confirmed the recombinant plasmid contained the complete VP(3) CDS sequence. Western blotting detected a 14.03 kD protein expression from the transfected cells, which was the expecting band of VP(3) gene. The growth of CNE-2 cells that expressed VP(3) gene was inhibited,while the growth of CNE-2 cells that did not express VP(3) gene was not inhibited. CONCLUSION: VP(3) gene can kill nasopharyngeal carcinoma cell CNE-2.
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OBJECTIVE: To investigate the killing effects of VP(3) on nasopharyngeal carcinoma cell line CNE-2. METHODS: Plasmid expression vector pcDNA3.1(-) CMV.VP(3)-His was constructed and identified by Kpn I/EcoR I endonuclease analysis, and then sequenced to verify successful insertion in the sense direction of VP(3) gene. pcDNA3.1(-) CMV.VP(3)-His and pcDNA3.1(-)-His expression plasmid was transiently transfected into nasopharyngeal carcinoma cell line CNE-2 . VP(3) protein expression was detected by Western blotting. MTT assay was used to determine the killing effects of VP(3) gene on nasopharyngeal carcinoma cell line CNE-2. RESULTS: Endonuclease analysis and sequencing confirmed the recombinant plasmid contained the complete VP(3) CDS sequence. Western blotting detected a 14.03 kD protein expression from the transfected cells, which was the expecting band of VP(3) gene. The growth of CNE-2 cells that expressed VP(3) gene was inhibited,while the growth of CNE-2 cells that did not express VP(3) gene was not inhibited. CONCLUSION: VP(3) gene can kill nasopharyngeal carcinoma cell CNE-2.
Key concepts: Nasopharyngeal carcinoma, Transfection, Molecular biology, Cell culture, Plasmid, Blot, Biology, Gene