As a de-ubiquitin enzyme, ubiquitin C-terminal hydrolase (UCH)-L1 has been shown to be overexpressed in several human cancers. However, the function of UCH-L1 in invasion of breast cancers is still unclear. Here we report that the expression of UCH-L1 is significantly higher in cancer cells with higher invasive ability. While ectopic UCH-L1 expression failed to alter cell proliferation in MCF-7 cells, it caused a significant upregulation of cellular invasion. Furthermore, siRNA mediated knockdown of UCH-L1 led to suppression of invasion in UCH-L1 overexpressing MCF-7 cells. In order to identify molecular mechanisms underlying these observations, a novel in vitro proximity-dependent biotin identification method was developed by fusing UCH-L1 protein with a bacterial biotin ligase (Escherichia coli BirA R118G, BioID). Streptavidin magnetic beads pulldown assay revealed that UCH-L1 can interact with Akt in MCF-7 cells. Pulldown assay with His tagged recombinant UCH-L1 protein and cell lysate from MCF-7 cells further demonstrated that UCH-L1 preferentially binds to Akt2 for Akt activation. Finally, we demonstrated that overexpression of UCH-L1 led to activation of Akt as evidenced by upregulation of phosphorylated Akt. Thus, these findings demonstrated that UCH-L1 promotes invasion of breast cancer cells and might serve as a potential therapeutic target for treatment of human patients with breast cancers.
|Number of pages||10|
|Journal||Journal of Cellular Biochemistry|
|State||Published - Jan 2018|
Bibliographical noteFunding Information:
American Heart Association, Grant number: 12SDG12070174; National Heart, Lung, and Blood Institute, Grant number: HL124122; National Cancer Institute, Grant number: CA109371; National Natural Science Foundation of China, Grant number: 81401155; National Institute of Arthritis and Musculoskeletal and Skin Diseases, Grant number: AR067766
This work was supported by U.S. National Institute of Health (NIH) Grant #CA109371 to J.Y., NIH grants #HL124122, #AR067766 and American Heart Association Grant 12SDG12070174 to H.Z., and the National Natural Science Foundation of China (Grant No. 81401155) to T.T.
© 2017 Wiley Periodicals, Inc.
- breast cancer
ASJC Scopus subject areas
- Molecular Biology
- Cell Biology