[Overexpression of CHIP in chronic myeloid leukemia K562 cells induces mitotic abnormality].
This study was aimed to investigate the possible influence of a novel E3 ubiquitin ligase CHIP (carboxyl terminus of Hsc70/Hsp70-interacting protein) on biological characteristics of cancer cells. Stable overexpression models in CML K562 cells were established via lipofectamine-mediated wild type CHIP and its TPR or U-box deletion mutants gene transfection. Followed G418 pressure selection, K562-CHIP stable transfected cell clones were obtained by limited dilution. The proliferation status and cell cycle were observed by MTT assay and FACS. The expression of related proteins and morphological changes were detected by Western blot and Wright-Giemsa staining. The results showed that overexpression of wild type CHIP did not inhibit cell proliferation, but slightly increased cell ratio of G(2)/M phase. CHIP gene had no effect on the stability of BCR-ABL kinase protein. HDAC inhibitor FK228-induced BCR-ABL degradation did not enhanced by CHIP. Notably the enlarged cells and abnormal mitotic cells remarkably increased in K562 WT-CHIP cells, indicating that CHIP may involve in the regulation of mitotic process. It is concluded that wild type CHIP induces mitotic abnormity in K562 cells.
Gao
Ying
Y
Department of Pathobiology, Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
Wang
Yan
Y
Zhang
Xu-Hui
XH
Chen
Guo-Zhu
GZ
DU
Zhi-Yan
ZY
Xu
Yuan-Ji
YJ
Yu
Xiao-Dan
XD
D006360:Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D020014:K562 Cells; D008938:Mitosis; D028861:Nuclear Pore Complex Proteins / Q000235:genetics / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D014162:Transfection; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*
D006360:Heat-Shock Proteins; D028861:Nuclear Pore Complex Proteins; D011518:Proto-Oncogene Proteins; C073597:TPR protein, human; C467728:STUB1 protein, human; D044767:Ubiquitin-Protein Ligases