[VP3对人鼻咽癌CNE-2细胞系的杀伤作用]

[Killing effect of VP3 on human nasopharyngeal carcinoma cell line CNE-2 cells].

作者信息

Xu Jing, Qiu Yuan-zheng, Tang Yao-yun, Tian Yong-quan, Xiao Xian-zhong, Zhao Su-ping

机构信息

Department of Otolaryngology, Xiangya Hospital, Central South University, Changsha 410008, China.

出版信息

Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2006 Oct;31(5):706-9.

PMID:17062935

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.

摘要

目的

研究VP(3)对鼻咽癌CNE - 2细胞系的杀伤作用。

方法

构建质粒表达载体pcDNA3.1(-)CMV.VP(3)-His，经Kpn I/EcoR I内切酶分析鉴定，然后测序以验证VP(3)基因正向成功插入。将pcDNA3.1(-)CMV.VP(3)-His和pcDNA3.1(-)-His表达质粒瞬时转染至鼻咽癌CNE - 2细胞系。通过蛋白质免疫印迹法检测VP(3)蛋白表达。采用MTT法测定VP(3)基因对鼻咽癌CNE - 2细胞系的杀伤作用。