EB病毒潜伏膜蛋白对人鼻咽癌细胞系CNE1生长分化的影响

Effectsof EBvirus LMPon the differentiation and growth of human nasopharyngeal carcinoma cellline CNE1

目的 研究ＥＢ病毒潜伏膜蛋白（ＥＢＶＬＭＰ） 对人高分化鼻咽癌细胞生长分化的影响。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ＥＢＶＬＭＰ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、流式细胞仪（ＦＣＭ）和裸鼠体内成瘤实验等，观察细胞生长分化的变化。结果 ＥＢＶＬＭＰ在体外可明显促进ＣＮＥ１ 细胞的增殖，实验组平均吸光度（ Ａ） 比值（３．９８ ±０ ．１１） 高于空白组（２．３６ ±０．０５） 及阴性对照组（２．７５ ±０．０７，Ｐ＜ ０．０１） ；实验组细胞软琼脂克隆形成率为２５ ．２％ （３７８／１ ５００ 个） ，显著高于空白组１１ ．２％（１６８／１ ５００ 个） 及阴性对照组１３ ．４ ％ （２０１／１ ５００ 个） ；ＦＣＭ 法测定细胞角蛋白表达， 实验组阳性率（８２．７ ％） 比空白组（９２．５ ％ ）及阴性对照组（９５．７％ ） 显著降低（Ｐ＜ ０．０５） ；ＣＮＥ１ 及转染细胞系裸小鼠移植结果表明，实验组瘤组织切片细胞明显变小， 部分呈梭形，异型性增大，有向低分化鳞癌发展倾向；移植瘤潜伏期（２６ ．６±７．７） 天、体内倍增时间（２ ．５１±０．１８）

Objective To study the effects of EBV LMPgene on differentiation and growth of human nasopharyngealcarcinoma (NPC) cell. Methods With NPCcellline (CNE1) astarget,electroporation wasused totransfect EBV LMPgene and the vector into CNE1 cells. The proliferation changes of transfectants were measuredin vitro by proliferationexperiment.FCMmethod andtransplantationinnude mice were used.Results LMPpromoted the growth ability of NPCCNE1 cellsin vitro.The average Aratio(3.98 ±0.11)inthe treatment group was higherthan inthe controlgroup(2.36 ±0.05)and alsointhe negative controlgroup(2 .75 ±0 .07 ,P< 0.01) .Cloneformingtest(25 .2% ,378/1 500)inthetreatmentgroup washigherthaninthecontrolgroups(11 . 2% ,168/1 500)and negative control groups(13.4% ,201/1 500) .When FCM method was used,the keratin positiverate (82 .7 % )inthetreatmentgroup was markedly decreased ascompared withthe control(92.5% ) and negative controlgroups(95.7% , P< 0 .05) .Latentperiod (26 .6±7.7)daysandinternaldoubletime(2.51 ±0.18)days oftransplant tumor ofthe treatment group were markedly shortened,tumortake rate(5/6) was increased,cells ofthetransplanttumorinthetreatmentgroup showed an image ofsmallsize,greatallotype,and atrend oflow differentiation.Conclusion The EBV LMPcan promotethegrowthandinhibitthe differentiation of NPCCNE1 cells.EBV LMPmayplay animportantroleinthe developmentofNPCandinstudyingits mechanism .