The deubiquitylase UCHL3 maintains cancer stem-like properties by stabilizing the aryl hydrocarbon receptor

Ling Chen,Min Wang,Yating Liu,Shuang Liu,Xiang Wang,Lingqiang Zhang,Desheng Xiao,Zuli Wang,Ying Shi,Yongguang Tao,Chao Mao,Ya Cao,Lianlian Ouyang,Na Liu,Yan Cheng,Bin Yan,Shouping Liu

doi:10.1038/s41392-020-0181-3

Abstract

Cancer stem cells (CSCs) exhibit highly aggressive and metastatic features and resistance to chemotherapy and radiotherapy. Aryl hydrocarbon receptor (AhR) expression varies among non-small cell lung cancers (NSCLCs), and the mechanisms that support abnormal AhR expression in CSCs remain elusive. Here, we identified ubiquitin carboxyl terminal hydrolase L3 (UCHL3), a DUB enzyme in the UCH protease family, as a bona fide deubiquitylase of the AhR in NSCLC. UCHL3 was shown to interact with, deubiquitylate, and stabilize AhR in a manner dependent on its deubiquitylation activity. Moreover, we showed that UCHL3 promotes the stem-like characteristics and potent tumorigenic capacity of NSCLC cells. UCHL3 increased AhR stability and the binding of AhR to the promoter regions of the “stemness” genes ATP-binding cassette subfamily G member 2 (ABCG2), KLF4, and c-Myc. Depletion of UCHL3 markedly downregulated the “stemness” genes ABCG2, KLF4, and c-Myc, leading to the loss of self-renewal and tumorigenesis in NSCLCs. Furthermore, the UCHL3 inhibitor TCID induced AhR degradation and exhibited significantly attenuated efficacy in NSCLC cells with stem cell-like properties. Additionally, UCHL3 was shown to indicate poor prognosis in patients with lung adenocarcinoma. In general, our results reveal that the UCHL3 deubiquitylase is pivotal for AhR protein stability and a potential target for NSCLC-targeted therapy.

Highlights

Proteins are decorated with a diverse array of posttranslational modifications (PTMs) that regulate their spatial and temporal functions
We observed that both Aryl hydrocarbon receptor (AhR) and reduced tumor size, volume, and weight (Fig. 3f–h); ubiquitin carboxyl terminal hydrolase L3 (UCHL3) protein levels were increased in lung cancer tissues there was no significant difference in body weight between the compared with normal tissues, and a positive correlation between two groups (Supplementary Fig. S3b)
AhR and UCHL3 plasmids were cotransfected into HEK293T cells for protein–protein interaction analysis via coimmunoprecipitation, and we discovered that the UCHL3 protein

Summary

1234567890();,: INTRODUCTION

Proteins are decorated with a diverse array of posttranslational modifications (PTMs) that regulate their spatial and temporal functions. Protein ubiquitination is a posttranslational modification that regulates all kinds of biological processes by influencing the stabilization, localization and function of substrate proteins.[1]. Using signaling pathway was shown to be related to radiation resistance lentivirus, UCHL3 was overexpressed in A549 and H1299 cells and and the stem-like characteristics of cancer cells, whereas AhR knocked down in H358 cells. Tissues with stem cell characteristics, have the ability to undergo UCHL3 overexpression in the A549 and H1299 cell lines self-renewal and the potential for nondirectional differentiation; obviously increased colony formation ability

DISCUSSION

Findings

MATERIALS AND METHODS