EB病毒潜伏膜蛋白对鼻咽癌细胞体内外增殖力的影响

Effects of EBV-LMP on Proliferation of Nasopharyngeal Carcinoma Cell Line CNE1

目的:研究EBVLMP对人高分化鼻咽癌细胞株CNE1体内外增殖能力的影响。方法:以人高分化鼻咽癌细胞株(CNE1)为对象,采用电穿孔基因转染技术,将重组EBVLMP表达质粒转染CNE1细胞。以载体质粒转染及CNE1细胞为对照,用细胞体外增殖实验、细胞软琼脂克隆形成率测定、流式细胞术、PCNA检测和裸鼠成瘤实验,观察细胞增殖能力的变化。结果:结晶紫染色法检测体外细胞增殖,结果表明,A比值实验组高于空白组及阴性对照组(P<001);实验组细胞软琼脂克隆形成率显著高于空白组及阴性对照组(P<001);FCM法测定细胞周期,实验组S期细胞比空白组及阴性对照组显著增高,S期细胞达349%;PCNA免疫组化染色,实验组细胞PCNA阳性率明显高于空白组及阴性对照组(P<001)。CNE1及转染细胞系裸小鼠移植结果表明,LMP表达细胞系移植瘤潜伏期、体内倍增时间明显缩短(P<001),成瘤率、瘤重显著增高(P<001/005)。结论:EBVLMP对CNE1细胞体内外增殖能力有明显的促进作用,提示LMP在NPC细胞演进过程中可能起重要作用,为进一步深入研究提供了依据。

Objective:The effect of human nasopharyngeal carcinoma cells (NPC) on proliferation transfected by EBV-LMP gene were studied.Methods:NPC cell line (CNE1) as target,Electroporation was used to transfect EBV-LMP gene and the vector into CNE1 cells.In vitro,the proliferation changes of transfectants were measured by proliferation experiment,clone forming,FCM method and PCNA test.Results:The proliferation experiment and clone forming results showed that,the LMP can contribute to increase the proliferation ability of NPC CNE1 cells,treatment group relative to control group and negative control group, P <0 01;FCM method test the cell cycle results show that,cells on S phase is 34 9% in the treatment group;The PCNA positive rate of treatment group cells sufficiently increased, P <0 01.The results of transplant of CNE1 and transfectant in nude mice showed:The latent period and internal doubling time of transplant tumor of LMP expressive cell line was sufficiently shorted, P <0 01,tumor formation rate and mass weight increased, P <0 01/0 05.Conclusion:The EBV-LMP can increase the proliferation ability of NPC CNE1 cells.The results suggests that EBV-LMP may have a important role in the development of NPC.