VP_3对鼻咽癌细胞CNE-2细胞株的体外杀伤效应  

Killing effect of VP_ 3 on human nasopharyngeal carcinoma cell line CNE-2 cells

作　　者：徐婧[1] 邱元正[1] 唐瑶云[1] 田勇泉[1] 肖献忠[2] 赵素萍[1] 

机构地区：[1]中南大学湘雅医院耳鼻咽喉科,长沙410078 [2]中南大学湘雅医学院病理生理学教研室,长沙410078

出　　处：《中南大学学报（医学版）》2006年第5期706-709,共4页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金(30300414)

摘　　要：目的:体外实验观察VP3对鼻咽癌CNE-2细胞株的杀伤效应。方法:构建质粒表达载体pcDNA3.1(-)CMV.VP3-His,KpnI/EcoRI酶切鉴定,测序分析VP3基因序列。脂质体法将pcDNA3.1(-)CMV.VP3-His和pcDNA3.1(-)-His分别瞬时转染鼻咽癌细胞CNE-2细胞株,Western印迹鉴定VP3基因的表达。四甲基偶氮唑盐(methylthiazoletetrazolium,MTT)法检测VP3对鼻咽癌细胞株CNE-2的杀伤效应。结果:酶切和测序分析证实重组质粒中包含完整的VP3基因的编码序列,Western印迹在转染细胞总蛋白中检测到该基因表达的16.03kD的蛋白。表达VP3基因的CNE-2细胞生长受到抑制,而不表达VP3基因的CNE-2细胞生长未受到抑制。结论:VP3可杀伤鼻咽癌细胞CNE-2细胞株。Objective To investigate the killing effects of VP3 on nasopharyngeal carcinoma cell line CNE-2. Methods Plasmid expression vector pcDNA3. 1 （ - ） CMV. VP3- His was constructed and identified by Kpn I/EcoR I endonuclease analysis, and then sequencd to verify successful insertion in the sense direction of VP3 gene. pcDNA3. 1 （ - ） CMV. VP3-His and pcDNA3. 1 （ - ） - His expression plasmid was transiently transfected into nasopharyngeal carcinoma cell line CNE-2 . VP3 protein expression was detected by Western blotting. MTT assay was used to determine the killing effects of VP3 gene on nasopharyngeal carcinoma cell line CNE-2. Results Endonuclease analysis and sequencing confirmed the recombinant plasmid contained the complete VP3 CDS sequence. Western blotting detected a 14.03 kD protein expression from the transfected cells, which was the expecting band of VP3 gene. The growth of CNE-2 cells that expressed VP3 gene was inhibited, while the growth of CNE-2 cells that did not express VP3 gene was not inhibited. Conclusion VP3 gene can kill nasopharyngeal carcinomacell CNE-2.

关 键 词：VP3 杀伤 鼻咽癌 

分 类 号：R739.63[医药卫生—肿瘤]