USP13 promotes breast cancer metastasis through FBXL14-induced Twist1 ubiquitination

Abstract

Purpose Epithelial-to-mesenchymal transition (EMT) is an important cause of high mortality in breast cancer. Twist1 is one of the EMT transcription factors (EMT-TFs) with a noticeably short half-life, which is regulated by proteasome degradation pathways. Recent studies have found that USP13 stabilizes several specific oncogenic proteins. As yet, however, the relationship between Twist1 and USP13 has not been investigated. Methods Co-Immunoprecipitation, GST-pulldown, Western blot, qRT-PCR and immunofluorescence assays were used to investigate the role of USP13 in de-ubiquitination of Twist1. Chromatin immunoprecipitation and Luciferase reporter assays were used to investigate the role of Twist1 in inhibiting USP13 reporter transcription. Scratch wound healing, cell migration and invasion assays, and a mouse lung metastases assay were used to investigate the roles of USP13 and Twist1 in promoting breast cancer metastasis. Results We found that Twist1 can be de-ubiquitinated by USP13. In addition, we found that the protein levels of Twist1 dose-dependently increased with USP13 overexpression, while USP13 knockdown resulted in a decreased expression of endogenous Twist1. We also found that USP13 can directly interact with Twist1 and specifically cleave the K48-linked polyubiquitin chains of Twist1 induced by FBXL14. We found that the effect of USP13 in promoting the migration and invasion capacities of breast cancer cells can at least partly be achieved through its regulation of Twist1, while Twist1 can inhibit the transcriptional activity of USP13. Conclusions Our data indicate that an interplay between Twist1 and USP13 can form a negative physiological feedback loop. Our findings show that USP13 may play an essential role in breast cancer metastasis by regulating Twist1 and, as such, provide a potential target for the clinical treatment of breast cancer.

Modulation of Epithelial–Mesenchymal Transition Is a Possible Underlying Mechanism for Inducing Chemoresistance in MIA PaCa-2 Cells against Gemcitabine and Paclitaxel

“…In fact, in addition to two key transcription factors, Snail and Twist, various signaling pathways could also be responsible factors such as Notch, tumor necrosis factor-alpha (TNFα), transforming growth factor beta (TGF-β), and hypoxia-inducible factor-1 alpha (HIF1α), which are involved in the induction of EMT in pancreatic cancer cells [65]. Quite interestingly, in the current study, all of the up-stream regulators and/or causal network master regulators estimated by the IPA analysis of the RNA sequencing, that is, STAT3 [42], GLI1 [43], ZNF367 [53], NKX3-2 [47], ZIC2 [51] IFIT2 [46], HEY1 [55] and FBXL14 [60] were identified as being directly or indirectly related to the EMT mechanisms of cancerous cells. Furthermore, among four factors, FABP5, IQSEC3, GASK1B, and SCN1A were identified as unique genes associated with their antisense RNA or pseudogene among the prominently altered DEGs (Log2 fold changes more than 6 or less than −6), FABP5 [66] and GASK1B [67] are also identified as modulators of cancerous EMT.…”

Roles of ubiquitin‑specific protease 13 in normal physiology and tumors (Review)

“…USP13 regulates DNA repair, senescence and proteasomal degradation via interactions with RAP80-BRCA1 complex, cell senescence related E3 ubiquitin ligase murine double minute 2 (MDM2) and inflammation regulation-associated protease inactive rhomboid protein 2 (IRHOM2), respectively, and via hydrolyzation of K63-linked ubiquitin chains ( 67 , 92 ). USP13 is also involved in the cleavage of K48-linked polyubiquitin chains in some cases, such as the upregulation of Twist1 ( 79 ). The regulatory function of USP13 in hydrolyzing different types of ubiquitin chains is complex and it may affect the tumor progression.…”

“…While the expression of Twist1 could be regulated by USP13, USP13 interacts with Twist1 and stabilizes its expression via the ubiquitin-proteasome pathway. High expression of USP13 promotes the migration and invasion of tumors such as breast cancer through Twist1 ( 79 ).…”