Exp Mol Med.  2014 May;46(5):e97. 10.1038/emm.2014.23.

Silencing of KIF14 interferes with cell cycle progression and cytokinesis by blocking the p27(Kip1) ubiquitination pathway in hepatocellular carcinoma

• Xu H
• Choe C
• Shin SH
• Park SW
• Kim HS
• Jung SH
• Yim SH
• Kim TM
• Chung YJ

• KMID: 2155362
• DOI: http://doi.org/10.1038/emm.2014.23

Abstract

Although it has been suggested that kinesin family member 14 (KIF14) has oncogenic potential in various cancers, including hepatocellular carcinoma (HCC), the molecular mechanism of this potential remains unknown. We aimed to elucidate the role of KIF14 in hepatocarcinogenesis by knocking down KIF14 in HCC cells that overexpressed KIF14. After KIF14 knockdown, changes in tumor cell growth, cell cycle and cytokinesis were examined. We also examined cell cycle regulatory molecules and upstream Skp1/Cul1/F-box (SCF) complex molecules. Knockdown of KIF14 resulted in suppression of cell proliferation and failure of cytokinesis, whereas KIF14 overexpression increased cell proliferation. In KIF14-silenced cells, the levels of cyclins E1, D1 and B1 were profoundly decreased compared with control cells. Of the cyclin-dependent kinase inhibitors, the p27Kip1 protein level specifically increased after KIF14 knockdown. The increase in p27Kip1 was not due to elevation of its mRNA level, but was due to inhibition of the proteasome-dependent degradation pathway. To explore the pathway upstream of this event, we measured the levels of SCF complex molecules, including Skp1, Skp2, Cul1, Roc1 and Cks1. The levels of Skp2 and its cofactor Cks1 decreased in the KIF14 knockdown cells where p27Kip1 accumulated. Overexpression of Skp2 in the KIF14 knockdown cells attenuated the failure of cytokinesis. On the basis of these results, we postulate that KIF14 knockdown downregulates the expression of Skp2 and Cks1, which target p27Kip1 for degradation by the 26S proteasome, leading to accumulation of p27Kip1. The downregulation of Skp2 and Cks1 also resulted in cytokinesis failure, which may inhibit tumor growth. To the best of our knowledge, this is the first report that has identified the molecular target and oncogenic effect of KIF14 in HCC.

Keyword

cell cycle; cytokinesis; hepatocellular carcinoma; KIF14; p27(kip1); ubiquitination

MeSH Terms

Carcinoma, Hepatocellular/*metabolism
Cyclin-Dependent Kinase Inhibitor p27/genetics/*metabolism
Cyclins/genetics/metabolism
*Cytokinesis
Gene Silencing
Hep G2 Cells
Humans
Kinesin/genetics/*metabolism
Liver Neoplasms/*metabolism
Oncogene Proteins/genetics/*metabolism
Proteasome Endopeptidase Complex/metabolism
RNA, Messenger/genetics/metabolism
S-Phase Kinase-Associated Proteins/genetics/metabolism
*Ubiquitination
Cyclins
Oncogene Proteins
RNA, Messenger
S-Phase Kinase-Associated Proteins
Cyclin-Dependent Kinase Inhibitor p27
Proteasome Endopeptidase Complex
Kinesin