β-catenin is regulated by USP9x and mediates resistance to TRAIL-induced apoptosis in breast cancer

Ouyang,Wen; Zhang,Shimin; Yang,Bo; Yang,Chunxu; Zhang,Junhong; Zhou,Fuxiang; Xie,Conghua

doi:10.3892/or.2015.4463

β-catenin is regulated by USP9x and mediates resistance to TRAIL-induced apoptosis in breast cancer

Authors:
- Wen Ouyang
- Shimin Zhang
- Bo Yang
- Chunxu Yang
- Junhong Zhang
- Fuxiang Zhou
- Conghua Xie
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Affiliations: Department of Radiation and Medical Oncology, Zhongnan Hospital, Wuhan University, Wuhan, Hubei 430071, P.R. China, Hubei Key Laboratory of Tumor Biological Behaviors, Zhongnan Hospital, Wuhan University, Wuhan, Hubei 430071, P.R. China
Published online on: November 27, 2015 https://doi.org/10.3892/or.2015.4463
Pages: 717-724

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Abstract

To investigate the regulatory mechanisms of decoy receptor expression in TRAIL-resistant breast cancer MCF-7 cells, cytotoxicity and apoptosis assays were applied to examine sensitivity to TRAIL in breast cancer cells. Immunofluorescence and immunoprecipitation were used to detect the co-localization and interaction of USP9x and β-catenin. Luciferase assay was used to examine activity of the DcR1/DcR2/OPG reporter. Overexpression/silencing of β-catenin was performed to confirm β-catenin mediated transcription of the decoy receptors. Additionally, silencing of USP9x was performed to prove that USP9X stabilizes β-catenin and mediates TRAIL-resistance. It was found that USP9x interacted with β-catenin and inhibited the degradation of β-catenin through the deubiquitination of β-catenin. Luciferase reporter assays showed induction of DcR1/DcR2/OPG reporter activity observed upon co-transfection of β-catenin and Tcf-4. The overexpression/silencing of β-catenin further confirmed the role of β-catenin in the regulation of transcription of the decoy receptors. Silencing of USP9x directly evidenced that USP9x affected the protein expression level of β-catenin, the transcription level of the decoy receptors, and reversed TRAIL-resistance of MCF-7 cells. In conclusion, USP9x interacted with and stabilized β-catenin through deubiquitination to mediate transcription of the decoy receptors in breast cancer cells. Our results offer new insights into the mechanisms of resistance to TRAIL, and USP9x could potentially be a therapeutic target for TRAIL-resistant breast cancers.

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