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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。
To investigate the tumorigenicity of different domains of Epstein-Barr virus LMP1 in nasopharyngeal carcinoma, and to provide experimental evidence for elucidating the molecular pathogenesis of NPC and finding molecular targets for the treatment of NPC. The transfected blank vector was used as a control. HNE2-LMP1 (1 ~ 815), HNE2-LMP1 (1 ~ 231) and HNE2-LMP1 △ were transfected into NPC cell lines stably expressing different LMP1 mutants by electroporation. 187 ~ 351, and using these cell lines as materials, the viable cells were detected by MTT assay and BrdU incorporation assay. The rate of soft agar colony formation and the ability of tumorigenesis in nude mice were compared to observe the expression of LMP1 Effect of different domains on the growth of nasopharyngeal carcinoma cells. LMP1 (1 ~ 231) and LMP1 △ 187 ~ 351 significantly promoted the proliferation of HNE2 cells in vitro. The average absorbance (A) ratio of HNE2-LMP1 (1 ~ 231) and HNE2-LMP1 △ 187 ~ 351, BrdU incorporation, The colony formation rate was higher than that of HNE2-pSG5 and HNE2 (P <0.01), but there was no significant difference between HNE2-LMP1 (1 ~ 187) and HNE2-pSG5 and HNE2. The tumorigenicity, doubling time and average tumor weight of HNE2-LMP1 △ 187 ~ 351 and HNE2-LMP1 (1 ~ 231) were significantly higher than those of HNE2-pSG5 nasopharyngeal carcinoma cell line, with significant difference P <0 05). However, there was no significant difference in latency, doubling time and mean tumor weight between HNE2-LMP1 (1 ~ 187), HNE2-pSG5 and HNE2 nasopharyngeal carcinoma cell lines (P> 0.05). EBV LMP1CTAR1 and CTAR2 significantly promote the growth of HNE2 cells, suggesting that Epstein-Barr virus LMP1 may play an important role in the development of nasopharyngeal carcinoma.