EB病毒LMP1 CTAR1、CTAR2的表达促使人鼻咽癌细胞HNE2增殖

The LMP1 CTAR1 or LMP1 CTAR2 of Ep stein-Barr Virus Promotes Prolifer ation of Human HNE2 Cell Line

探讨EB病毒LMP1不同结构域在鼻咽癌中的致瘤作用,为阐明鼻咽癌分子发病机理,寻找治疗鼻咽癌的分子靶提供实验依据。以转染空白载体为对照,利用电穿孔转染方法,建立稳定表达LMP1不同突变体的鼻咽癌细胞系HNE2-LMP1(1～815)、HNE2-LMP1(1～231)、HNE2-LMP1△187～351,并以这些细胞系为材料,用MTT法检测增殖期活细胞,BrdU掺入法检测细胞增殖状况,比较各组细胞的软琼脂集落形成率和裸鼠成瘤能力,以观察LMP1不同的结构域对鼻咽癌细胞生长的影响。LMP1(1～231)和LMP1△187～351在体外明显促进HNE2细胞增殖,HNE2-LMP1(1～231)、HNE2-LMP1△187～351平均吸光度(A)比值、BrdU掺入率、软琼脂集落形成率均高于HNE2-pSG5与HNE2(P<0 01),而HNE2-LMP1(1～187)与HNE2-pSG5、HNE2相比,这些指标无明显差别。HNE2-LMP1△187～351和HNE2-LMP1(1～231)的裸鼠成瘤潜伏期、倍增时间与平均瘤重明显高于HNE2-pSG5鼻咽癌细胞系,其差异有显著的统计学意义(P<0 05)。而HNE2-LMP1(1～187)、HNE2-pSG5和HNE2鼻咽癌细胞系在潜伏期、倍增时间与平均瘤重方面两两比较,差异无显著的统计学意义(P>0 05)。EB病毒LMP1CTAR1和CTAR2对HNE2细胞生长有明显促进作用,提示EB病毒LMP1可能在鼻咽癌的发生发展中起着重要的作用。

This paper was to investi gate the carcinogenic effect of differe nt regions of LMP1 gene of Epstein-Ba rr virus on proliferation of human nasopharyngeal carcinoma cell line HNE2The stable transfectant cell lines were established by introduc ing LPM1 cDNA,LMP1(1-185),LMP1(1-2 31)(LMP1 CTAR1),LMP1△187-351(LMP1 CTAR2) and the vector (pSG5) into HNE2-an EBV-negative cell line der ived from poorly differentiated na sopharyngeal carcinoma by electro porotionUsing these cell lines as models,the proliferative changes o f transfectants were measured in v itro by proliferative experiment,B rdU incorporation ratio assay,clon ing forming assay and transplantat ion in nude miceThe results showe d that the LMP1(1-231) and LMP1△187 -351 promoted the growth ability o f NPC HNE2 cells in vitroThe avera ge A ratio,BrdU incorporation rati o,cloning forming ability of the H NE2-LMP1(1-231) and HNE2-LMP1△187 -351 cells were higher than those of HNE2-pSG5 cell and HNE2 (P<001)Incubation period and i nternal doubling time of transplanted tumor of HNE2-LMP1,HNE2-LMP1(1-231 ) and HNE2-LMP1△187-351 were marke dly shortenedIt suggests that LMP1 (1-231) and LMP1△187-351 can promo te the growth of NPC HNE2 cells,an d the LMP1 possibly plays an impor tant role in the development of NP C