LMP1经TRADD促进鼻咽癌SP18细胞增殖

摘要

目的: 探讨潜伏性膜蛋白1(LMP1)羧基末端活化区2(CTAR2)的肿瘤坏死因子受体相关死亡结构域(TRADD)在鼻咽癌SP18细胞增殖中的作用及机制.方法: 首先建立表达LMP1及CTAR2突变型LMP1(LMP1TRADD)的SP18细胞系.其次采用细胞生长曲线、平皿集落形成实验、软琼脂集落实验和流式细胞术观察LMP1TRADD对SP18细胞增殖的影响.然后选用基因芯片检测SP18-LMP1和SP18-LMP1TRADD细胞间的差异表达基因.结果: 细胞生长曲线、软琼脂集落和平皿集落形成实验结果均显示,SP18-LMP1细胞生长与集落形成能力均较SP18-LMP1TRADD细胞强(P<0.01).流式细胞术检测结果显示,SP18-LMP1细胞的增殖指数较SP18-LMP1TRADD细胞高(P<0.01).筛选出63个增殖相关基因,其中SP18-LMP1TRADD细胞中上调的基因33个,下调的基因30个.结论: TRADD活性区是LMP1促进SP18细胞增殖的重要功能活性位点.LMP1可能通过TRADD提高细胞增殖指数,诱导SP18细胞增殖.%AIM: To investigate the effects of TNF receptor-associated death domains (TRADD) of latent membrane protein 1 (LMP1) on the proliferation of nasopharyngeal cancer SP18 cells.METHODS: The SP18-LMP1 cells and SP18-LMP1TRADD cells, which expressed LMP1 and LMP1 TRADD proteins, respectively, were established.The proliferation of SP18 cells affected by LMP1TRADD was detected by cell counting to analyze the cell growth curve, and by colony formation assay, soft agar formation assay, and flow cytometry.Moreover, the expression profile of differential genes between SP18-LMP1 cells and SP18-LMP1TRADD cells was analyzed by gene chips.RESULTS: The cell growth curve, and the results of colony formation and soft agar formation displayed that the growth velocity and colony forming ability of SP18-LMP1 cells were stronger than those of SP18-LMP1TRADD cells (P<0.01).The results of flow cytometry analysis showed that the proliferation index of SP18-LMP1 cells was higher than that of SP18-LMP1TRADD cells (P<0.01).Sixty-three differentially expressed genes associated with cell proliferation were screened out, in which 33 genes were up-regulated and 30 were down-regulated in the SP18-LMP1TRADD cells.CONCLUSION: TRADD active region is an important functional site of LMP1 to promote the proliferation of SP18 cells.LMP1 may improve the cell proliferation index and induce the proliferation of SP18 cells through TRADD.