The key laboratory of laboratory medical diagnostics, ministry of education; the faculty of laboratory medicine, Chongqing Medical University, Chongqing PR China.
J Exp Clin Cancer Res. 2009 Dec 18;28(1):156. doi: 10.1186/1756-9966-28-156.
CENP-E, one of spindle checkpoint proteins, plays a crucial role in the function of spindle checkpoint. Once CENP-E expression was interrupted, the chromosomes can not separate procedurally, and may result in aneuploidy which is a hallmark of most solid cancers, such as hepatocellular carcinoma (HCC). We investigate the expression of CENP-E in human hepatocellular carcinoma,. and analyze the effect of low CENP-E expression on chromosome separation in normal liver cell line (LO2).
We determined its levels in HCC and para-cancerous tissues, human hepatocellular carcinoma-derived cell line (HepG2) and LO2 cell line using real time quantitative PCR (QPCR) and Western blot. Further to know whether reduction in CENP-E expression impairs chromosomes separation in LO2 cells. we knocked down CENP-E using shRNA expressing vector and then count the aneuploid in LO2 cells using chromosomal counts assay.
We found that both CENP-E mRNA and protein levels were significantly reduced in HCC tissues and HepG2 cells compared with para-cancerous tissues and LO2 cells, respectively. A significantly-increased proportion of aneuploid in these down-knocked LO2 cells compared with those treated with control shRNA vector.
Together with other results, these results reveal that CENP-E expression was reduced in human HCC tissue, and low CENP-E expression result in aneuploidy in LO2 cells.
作为纺锤体检验点蛋白之一的 CENP-E 在纺锤体检验点功能中起着至关重要的作用。一旦 CENP-E 的表达被中断,染色体就不能有序分离,可能导致非整倍体,这是非大多数实体瘤(如肝癌)的标志之一。我们研究了 CENP-E 在人肝癌中的表达,并分析了低 CENP-E 表达对正常肝细胞系(LO2)中染色体分离的影响。
我们使用实时定量 PCR(QPCR)和 Western blot 技术测定 HCC 和癌旁组织、人肝癌衍生细胞系(HepG2)和 LO2 细胞系中 CENP-E 的水平。为了进一步了解 CENP-E 表达降低是否会损害 LO2 细胞中的染色体分离,我们使用 shRNA 表达载体敲低 CENP-E,然后使用染色体计数法计算 LO2 细胞中的非整倍体。
与癌旁组织和 LO2 细胞相比,我们发现 HCC 组织和 HepG2 细胞中的 CENP-E mRNA 和蛋白水平均显著降低。与用对照 shRNA 载体处理的细胞相比,这些下调的 LO2 细胞中非整倍体的比例显著增加。
结合其他结果,这些结果表明 CENP-E 在人肝癌组织中表达降低,低 CENP-E 表达导致 LO2 细胞中的非整倍体。