The scaffold protein DLG4 facilitates RNF63-mediated ubiquitination and degradation of STAT3 in non-small cell lung cancer

Abstract Background The Disks-large homolog (DLG) family has been found to govern multiple key processes in human cancers. However, their role in non-small cell lung cancer (NSCLC) remains unknown. Methods The expression of DLG4 was determined by immunoblotting and q-PCR. The interacting proteins of...

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Main Authors: Shisong Chen, Hongjie Xu, Ning Li, Yang Yang, Ruxi Pang, Shuwei Zhang, Junjie Qiao, Hao Chen
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Cell Communication and Signaling
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Online Access:https://doi.org/10.1186/s12964-025-02334-5
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author Shisong Chen
Hongjie Xu
Ning Li
Yang Yang
Ruxi Pang
Shuwei Zhang
Junjie Qiao
Hao Chen
author_facet Shisong Chen
Hongjie Xu
Ning Li
Yang Yang
Ruxi Pang
Shuwei Zhang
Junjie Qiao
Hao Chen
author_sort Shisong Chen
collection DOAJ
description Abstract Background The Disks-large homolog (DLG) family has been found to govern multiple key processes in human cancers. However, their role in non-small cell lung cancer (NSCLC) remains unknown. Methods The expression of DLG4 was determined by immunoblotting and q-PCR. The interacting proteins of DLG4 were identified by affinity purification mass spectrometry. The ubiquitination level of STAT3 was verified by denaturation-IP. The protein interactions were determined by co-IP. The clinical significance of DLG4, RNF63, and STAT3 was evaluated by immunohistochemical staining. Results In this study, by evaluating the expression levels of human DLG protein (DLG1-DLG5), we found that DLG4 is significantly downregulated in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC), two major types of NSCLC. DLG4 overexpression impairs cell proliferation and epithelial-mesenchymal transition migration (EMT) of NSCLC cells. The xenograft model also verifies the inhibitory effects of DLG4 on tumor growth in vivo. Moreover, we determined that DLG4 functions as a novel regulator of STAT3. Mechanistically, DLG4 directly interacts with STAT3 and recruits E3 ubiquitin ligase RNF63 (MKRN3) to STAT3, which promotes STAT3 K48-linked polyubiquitination and proteasome-mediated degradation. Importantly, in human NSCLC specimens, endogenous DLG4 and RNF63 expression levels are inversely correlated with that of STAT3. Moreover, low DLG4 and RNF63 expression correlates with poor patient survival in NSCLC. Conclusion our findings define the role of DLG4 that can diminish NSCLC cell proliferation and tumorigenesis through degrading STAT3 in an RNF63-dependent manner. This work suggests a new treatment strategy against NSCLC caused by aberrant activation of STAT3.
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spelling doaj-art-6ed1d9aa631b47a0acf338e97229cea02025-08-20T04:03:07ZengBMCCell Communication and Signaling1478-811X2025-07-0123111510.1186/s12964-025-02334-5The scaffold protein DLG4 facilitates RNF63-mediated ubiquitination and degradation of STAT3 in non-small cell lung cancerShisong Chen0Hongjie Xu1Ning Li2Yang Yang3Ruxi Pang4Shuwei Zhang5Junjie Qiao6Hao Chen7Department of Cardiovascular Surgery, Shanghai General Hospital, Shanghai Jiao Tong UniversityDepartment of Cardiovascular Surgery, Shanghai Institute of Thoracic Cardiac Surgery, Changhai HospitalDepartment of Cardiothoracic Surgery, Naval Medical Center of PLA, Naval Medical UniversityDepartment of General Surgery, Chongming Hospital, Shanghai University of Medicine and Health SciencesDepartment of Gastroenterology, Shanghai Institute of Pancreatic Diseases, Changhai HospitalDepartment of Cardiovascular Surgery, Shanghai Institute of Thoracic Cardiac Surgery, Changhai HospitalDepartment of Cardiovascular Surgery, Shanghai Institute of Thoracic Cardiac Surgery, Changhai HospitalDepartment of Orthopaedics, Chongming Hospital, Shanghai University of Medicine and Health SciencesAbstract Background The Disks-large homolog (DLG) family has been found to govern multiple key processes in human cancers. However, their role in non-small cell lung cancer (NSCLC) remains unknown. Methods The expression of DLG4 was determined by immunoblotting and q-PCR. The interacting proteins of DLG4 were identified by affinity purification mass spectrometry. The ubiquitination level of STAT3 was verified by denaturation-IP. The protein interactions were determined by co-IP. The clinical significance of DLG4, RNF63, and STAT3 was evaluated by immunohistochemical staining. Results In this study, by evaluating the expression levels of human DLG protein (DLG1-DLG5), we found that DLG4 is significantly downregulated in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC), two major types of NSCLC. DLG4 overexpression impairs cell proliferation and epithelial-mesenchymal transition migration (EMT) of NSCLC cells. The xenograft model also verifies the inhibitory effects of DLG4 on tumor growth in vivo. Moreover, we determined that DLG4 functions as a novel regulator of STAT3. Mechanistically, DLG4 directly interacts with STAT3 and recruits E3 ubiquitin ligase RNF63 (MKRN3) to STAT3, which promotes STAT3 K48-linked polyubiquitination and proteasome-mediated degradation. Importantly, in human NSCLC specimens, endogenous DLG4 and RNF63 expression levels are inversely correlated with that of STAT3. Moreover, low DLG4 and RNF63 expression correlates with poor patient survival in NSCLC. Conclusion our findings define the role of DLG4 that can diminish NSCLC cell proliferation and tumorigenesis through degrading STAT3 in an RNF63-dependent manner. This work suggests a new treatment strategy against NSCLC caused by aberrant activation of STAT3.https://doi.org/10.1186/s12964-025-02334-5NSCLCDLG4RNF63STAT3
spellingShingle Shisong Chen
Hongjie Xu
Ning Li
Yang Yang
Ruxi Pang
Shuwei Zhang
Junjie Qiao
Hao Chen
The scaffold protein DLG4 facilitates RNF63-mediated ubiquitination and degradation of STAT3 in non-small cell lung cancer
Cell Communication and Signaling
NSCLC
DLG4
RNF63
STAT3
title The scaffold protein DLG4 facilitates RNF63-mediated ubiquitination and degradation of STAT3 in non-small cell lung cancer
title_full The scaffold protein DLG4 facilitates RNF63-mediated ubiquitination and degradation of STAT3 in non-small cell lung cancer
title_fullStr The scaffold protein DLG4 facilitates RNF63-mediated ubiquitination and degradation of STAT3 in non-small cell lung cancer
title_full_unstemmed The scaffold protein DLG4 facilitates RNF63-mediated ubiquitination and degradation of STAT3 in non-small cell lung cancer
title_short The scaffold protein DLG4 facilitates RNF63-mediated ubiquitination and degradation of STAT3 in non-small cell lung cancer
title_sort scaffold protein dlg4 facilitates rnf63 mediated ubiquitination and degradation of stat3 in non small cell lung cancer
topic NSCLC
DLG4
RNF63
STAT3
url https://doi.org/10.1186/s12964-025-02334-5
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