电穿孔技术在EBV-LMP表达质粒转染人鼻咽癌细胞中的应用

APPLICATION OF ELECTROPORATION TO TRANSFECTING OF EBV-LMP EXPRESSING PLASMID INTO NPC CNE1 CELLS IN VITRO

目的：观察人鼻咽癌细胞用电穿孔技术进行体外基因转染所需的转染电击条件及EBV－LMP基因的转染表达。方法：以人高分化鼻咽癌细胞株（CNE1）为对象，用电穿孔基因转染技术，观察不同电压及电容条件电击癌细胞对其存活率的影响；将重组EBV－LMP表达质粒转染CNE1细胞，以载体质粒转染及未经转染的CNE1细胞为对照。结果：CNE1癌细胞存活率与电容及电压值的大小成反比。CNE1细胞转染较合适条件是电压值600～1200V／cm与电容250～25μF，癌细胞存活率为56.1％～70.2％。将重组EBV－LMP表达质粒转染CNE1细胞，获得LMP稳定高表达克隆。结论：电穿孔基因转染技术可用于人高分化鼻咽癌细胞（CNE1）进行基因转染。采用该技术建立了表达EBV－LMP的人鼻咽癌细胞模型。

Objective: To study the condition of electroporation on human nasopharyngeal carcinoma cells (NPC)and to get a cell model of stable transfection by EBV-LMP gene. Methods:Electroporation was done by using NPC cell line(CNE1 )as target at different condition. With the suitable condition,recombinant EBV-LMP gene and the vector were trans fected into CNE1 cells. Results: The survival rate of transfected CNE1 was inversely proportional to the values of capacitance and voltage which were used.The appropriate condition for transfection of CNE1, at which the transfectants could survive at 56.1 %~ 70. 2%,was 250~ 25μF and 600~ 1200V/cm. The stable high expressive LMP clone was gained by means of transfection of recombinant EBV-LMP gene and vector into CNE1 cells. Conclusiou:Electro poration might be a efficient method for trans fecting gene into CNE1 cells. The cell model of expressing EBV-LMP will contribute to study of NPC.