Abstract
Downregulation of the cylindromatosis (CYLD) tumor suppressor has been associated with breast cancer development and progression. Here, we report a critical role for CYLD in maintaining the phenotype of mammary epithelial cells in vitro and in vivo. CYLD downregulation or inactivation induced an epithelial to mesenchymal transition of mammary epithelial cells that was dependent on the concomitant activation of the transcription factors Yes-associated protein (YAP)/transcriptional coactivator with PDZ-binding motif (TAZ) and transforming growth factor beta (TGFβ)signaling. CYLD inactivation enhanced the nuclear localization of YAP/TAZ and the phosphorylation of Small Mothers Against Decapentaplegic (SMAD)2/3 proteins in confluent cell culture conditions. Consistent with these findings were the hyperplastic alterations of CYLD-deficient mouse mammary epithelia, which were associated with enhanced nuclear expression of the YAP/TAZ transcription factors. Furthermore, in human breast cancer samples, downregulation of CYLD expression correlates with enhanced YAP/TAZ-regulated target gene expression. Our results identify CYLD as a critical regulator of a signaling node that prevents the coordinated activation of YAP/TAZ and the TGFβ pathway in mammary epithelial cells, in order to maintain their phenotypic identity and homeostasis. Consequently, they provide a novel conceptual framework that supports and explains a causal implication of deficient CYLD expression in aggressive human breast cancers.
Highlights
The epithelial to mesenchymal transition (EMT) has been recognized as an important mechanism that can promote invasion and metastasis of breast cancer cells
In mammary epithelium, we used CRISPR/Cas9 to generate clones of non-transformed MCF10A cells that possess different mutations in the CYLD gene, which result in the loss of full-length CYLD protein (Figure 1a) and reduction of CYLD mRNA expression (Figure 1b) that is likely due to non-sense mRNA decay (NMD)
3c,d), a phenotype that was not observed in the cultures of invading into the surrounding extracellular matrix (Figure 3c,d), a phenotype that was not observed control cells. These findings indicate that loss of functional in mammary epithelial cells confers in the cultures of control cells. These findings indicate that loss of functional CYLD in mammary stem-like invasive are consistent with the aggressive behavior of CYLD-deficient epithelialand cells confersproperties, stem-like which and invasive properties, which are consistent with the aggressive human breast cancers
Summary
The epithelial to mesenchymal transition (EMT) has been recognized as an important mechanism that can promote invasion and metastasis of breast cancer cells (reviewed in [1]). The deubiquitinating domain of CYLD is located at the carboxyl-terminal region of the protein and three CAP-Gly domains are found within the CYLD amino terminal region, two of which are capable of interacting with microtubules and their associated proteins end-binding protein 1 (EB1) and histone deacetylase 6 (HDAC6) [20]. These interactions have been implicated in the regulation of microtubule dynamics by CYLD, which affect cell migration and various aspects of the cell cycle [21,22,23]. Cas and gRNAs that target gene (a) MCF10A cells infected were infected with lentiviral vectors expressing.
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
