纺锤体检查点蛋白Cenp-E过低表达与肝癌细胞染色体异常的关系  

Relation of low expression of spindle checkpoint protein CENP-E and chromosome abnormality in human hepatoma cells

作　　者：蔡勇 彭爱红[2] 

机构地区：[1]湖南博雅眼科医院检验科,湖南长沙410007 [2]湖南省临床检验中心,湖南长沙410007

出　　处：《湖南师范大学学报（医学版）》2011年第4期19-24,共6页Journal of Hunan Normal University(Medical Sciences)

摘　　要：目的:探讨纺锤体检查点蛋白E(Cenp-E)基因在肝癌细胞中的定位和作用,为进一步阐明肝癌细胞染色体数目异常的机理奠定基础。方法:利用FQ-PCR检测Cenp-E基因在用Nocodazole(诺考达唑)同时处理HepG-2细胞和LO2细胞中mRNA的表达水平。用间接免疫荧光技术观察Cenp-E蛋白在用Nocodazole(诺考达唑)同时处理HepG-2细胞和LO2细胞中的定位及表达。在LO2细胞中用RNAi技术干扰Cenp-E基因后,用FQ-PCR检测Cenp-E基因在干扰前后mRNA表达水平。并利用间接免疫荧光进一步评价干扰后的细胞功能情况。结果:Nocodazole处理前Cenp-E基因和蛋白在LO2和HepG-2细胞中表达无显著差异,处理后Cenp-E基因和蛋白在两种细胞表达都增高,但LO2细胞上调水平大于HepG-2细胞,差异具有统计学意义。间接免疫荧光结果显示Cenp-E蛋白主要定位细胞核内,并且在出现异常分裂的细胞核内Cenp-E蛋白的表达明显低于正常分裂的细胞核。在干扰Cenp-E后,间接免疫荧光显示干扰后的LO2细胞中出现核异常比例明显高于正常细胞。结论:Cenp-E过低表达可能是肝癌细胞染色体数目异常重要原因之一。Objective To study expression of Cenp-E localization and role in hepatoma cells by indirect immunofluorescence and RNAi.It imply important information for exploring the mechanism of chromosome numerical abnormality in human hepatoma cells.Methods We detected the mRNA of Cenp-E in the HepG-2 cells and LO2 cells with Nocodazole at the same time to synchronize them at mitosis before and after treating with Nocodazole by FQ-PCR.We observed the localization and expression of Cenp-E protein in the two kinds of cells by indirect immunofluorescence.After Cenp-E was interfered in LO2 cells by RNAi,we detected the mRNA expression of Cenp-E before and after interference by FQ-PCR,and then,further evaluated the cells,function after interference by indirect immunofluorescence.Results The results of FQ-PCR showed that the expression of Cenp-E in the two kinds of cells before treating with Nocodazole was no significant difference,the up-regulated expression level of BubR1 and Cenp-E in hepatic cell line L02 was higher than that in human hepatoma carcinoma cell line HepG2 during cell division phase.Indirect immunofluorescence showed that Cenp-E protein mainly located in the nucleus,and that the expression of Cenp-E protein in the nucleus with abnormality mitosis was significantly lower than that in the nucleus with normal mitosis.AfterCenp-E was interfered.Indirect immunofluorescence showed the ratio of dyskaryosis in the cells with interfered Cenp-E was significantly higher than that in the normal cells.Conclusion The low expression of Cenp-E may be an important reason of chromosome numerical abnormality in human hepatoma cells.

关 键 词：肝癌 纺锤体检查点 CENP-E 染色体数目异常 

分 类 号：R735.7[医药卫生—肿瘤]