Reduced expression of cenp-e in human hepatocellular carcinoma.
BACKGROUND
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).


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Liu
Zijie
Z
The key laboratory of laboratory medical diagnostics, ministry of education; the faculty of laboratory medicine, Chongqing Medical University, Chongqing PR China. redjacky502@yahoo.com.cn
Ling
Kang
K
Wu
Xia
X
Cao
Ju
J
Liu
Bin
B
Li
Suyan
S
Si
Qiong
Q
Cai
Yan
Y
Yan
Chen
C
Zhang
Yan
Y
Weng
Yaguang
Y
D000782:Aneuploidy; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D002868:Chromosomal Proteins, Non-Histone / Q000096:biosynthesis*; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D002868:Chromosomal Proteins, Non-Histone; D012333:RNA, Messenger; C068688:centromere protein E