Emodin promotes GSK-3β-mediated PD-L1 proteasomal degradation and enhances anti-tumor immunity in hepatocellular carcinoma

Abstract Background Programmed death-ligand 1 (PD-L1), a prominent immune checkpoint, interacts with programmed death protein-1 (PD-1) on cytotoxic T cells within tumors and promotes immune evasion. Emodin, which is known to destabilize PD-L1 in breast cancer, has great potential for enhancing anti-...

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Main Authors: Xuemei Yang, Weiguang Chen, Haitao Sun, Weicong Chen, Wei Xu, Chunyu He, Yang Liu, Ying Kuang, Yanhao Ma, Binglian Zhong, Chaojie Li, Guohuan Li, Qingfeng Du, Songqi He
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Language:English
Published: BMC 2025-08-01
Series:Chinese Medicine
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Online Access:https://doi.org/10.1186/s13020-025-01146-6
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author Xuemei Yang
Weiguang Chen
Haitao Sun
Weicong Chen
Wei Xu
Chunyu He
Yang Liu
Ying Kuang
Yanhao Ma
Binglian Zhong
Chaojie Li
Guohuan Li
Qingfeng Du
Songqi He
author_facet Xuemei Yang
Weiguang Chen
Haitao Sun
Weicong Chen
Wei Xu
Chunyu He
Yang Liu
Ying Kuang
Yanhao Ma
Binglian Zhong
Chaojie Li
Guohuan Li
Qingfeng Du
Songqi He
author_sort Xuemei Yang
collection DOAJ
description Abstract Background Programmed death-ligand 1 (PD-L1), a prominent immune checkpoint, interacts with programmed death protein-1 (PD-1) on cytotoxic T cells within tumors and promotes immune evasion. Emodin, which is known to destabilize PD-L1 in breast cancer, has great potential for enhancing anti-tumor immunity. However, whether emodin can modulate PD-L1 levels in hepatocellular carcinoma (HCC) and enhance anti-tumor immune response remains unclear. Materials and methods PD-L1 levels were assessed by western blot and RT-qPCR, the degradation mechanism was analyzed using specific inhibitors. Network pharmacology, molecular docking, and glycogen synthase kinase-3 beta (GSK-3β) modulation analyzes were performed to validate emodin’s target. In vivo anti-tumor effects were evaluated in H22 subcutaneous tumor model, and CD8+ T cells and RNA-seq data were analyzed. The synergistic effects of emodin and an anti-PD-L1 antibody were assessed. Results Emodin effectively reduced PD-L1 levels in H22 cells and increased anti-tumor activity in an H22 subcutaneous tumor model by promoting CD8+ T cells infiltration and TNF-α, IFN-γ, and granzyme B secretion. Mechanistically, emodin accelerated PD-L1 degradation through the proteasome pathway in both mouse and human HCC cell lines, as confirmed by the use of proteasome, lysosome and autophagy inhibitors. Network pharmacology analysis and molecular docking revealed that GSK-3β, a key regulator of PD-L1 degradation, is a target of emodin. Selective inhibitor-mediated suppression of GSK-3β largely reversed the regulatory effect of emodin on PD-L1. In contrast, overexpression of GSK-3β with a plasmid decreased PD-L1 protein levels and augmented emodin’s effect on PD-L1. Additionally, RNA-sequencing revealed the role of emodin in improving the immune responses in the tumor microenvironment. Finally, we observed a synergistic effect when the H22 cell subcutaneous tumor model was treated with emodin and anti-PD-L1 antibody. Conclusion Emodin exerts anti-tumor effects by promoting GSK-3β-mediated PD-L1 proteasomal degradation and enhancing the anti-tumor effects of CD8+ T cells, indicating that emodin may be a promising therapeutic option for HCC. Graphical Abstract
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spelling doaj-art-8f323e7a5ea54723bf07f8f6c20bb7672025-08-20T03:43:30ZengBMCChinese Medicine1749-85462025-08-0120111710.1186/s13020-025-01146-6Emodin promotes GSK-3β-mediated PD-L1 proteasomal degradation and enhances anti-tumor immunity in hepatocellular carcinomaXuemei Yang0Weiguang Chen1Haitao Sun2Weicong Chen3Wei Xu4Chunyu He5Yang Liu6Ying Kuang7Yanhao Ma8Binglian Zhong9Chaojie Li10Guohuan Li11Qingfeng Du12Songqi He13Affiliated Zhuhai Hospital, Southern Medical University (Zhuhai Hospital of Integrated Traditional Chinese and Western Medicine)School of Traditional Chinese Medicine, Southern Medical UniversitySchool of Traditional Chinese Medicine, Southern Medical UniversityAffiliated Zhuhai Hospital, Southern Medical University (Zhuhai Hospital of Integrated Traditional Chinese and Western Medicine)School of Traditional Chinese Medicine, Southern Medical UniversitySchool of Traditional Chinese Medicine, Southern Medical UniversitySchool of Traditional Chinese Medicine, Southern Medical UniversitySchool of Traditional Chinese Medicine, Southern Medical UniversitySchool of Traditional Chinese Medicine, Southern Medical UniversitySchool of Traditional Chinese Medicine, Southern Medical UniversityThe People’s Hospital Medical Group of XiangzhouAffiliated Zhuhai Hospital, Southern Medical University (Zhuhai Hospital of Integrated Traditional Chinese and Western Medicine)School of Traditional Chinese Medicine, Southern Medical UniversityAffiliated Zhuhai Hospital, Southern Medical University (Zhuhai Hospital of Integrated Traditional Chinese and Western Medicine)Abstract Background Programmed death-ligand 1 (PD-L1), a prominent immune checkpoint, interacts with programmed death protein-1 (PD-1) on cytotoxic T cells within tumors and promotes immune evasion. Emodin, which is known to destabilize PD-L1 in breast cancer, has great potential for enhancing anti-tumor immunity. However, whether emodin can modulate PD-L1 levels in hepatocellular carcinoma (HCC) and enhance anti-tumor immune response remains unclear. Materials and methods PD-L1 levels were assessed by western blot and RT-qPCR, the degradation mechanism was analyzed using specific inhibitors. Network pharmacology, molecular docking, and glycogen synthase kinase-3 beta (GSK-3β) modulation analyzes were performed to validate emodin’s target. In vivo anti-tumor effects were evaluated in H22 subcutaneous tumor model, and CD8+ T cells and RNA-seq data were analyzed. The synergistic effects of emodin and an anti-PD-L1 antibody were assessed. Results Emodin effectively reduced PD-L1 levels in H22 cells and increased anti-tumor activity in an H22 subcutaneous tumor model by promoting CD8+ T cells infiltration and TNF-α, IFN-γ, and granzyme B secretion. Mechanistically, emodin accelerated PD-L1 degradation through the proteasome pathway in both mouse and human HCC cell lines, as confirmed by the use of proteasome, lysosome and autophagy inhibitors. Network pharmacology analysis and molecular docking revealed that GSK-3β, a key regulator of PD-L1 degradation, is a target of emodin. Selective inhibitor-mediated suppression of GSK-3β largely reversed the regulatory effect of emodin on PD-L1. In contrast, overexpression of GSK-3β with a plasmid decreased PD-L1 protein levels and augmented emodin’s effect on PD-L1. Additionally, RNA-sequencing revealed the role of emodin in improving the immune responses in the tumor microenvironment. Finally, we observed a synergistic effect when the H22 cell subcutaneous tumor model was treated with emodin and anti-PD-L1 antibody. Conclusion Emodin exerts anti-tumor effects by promoting GSK-3β-mediated PD-L1 proteasomal degradation and enhancing the anti-tumor effects of CD8+ T cells, indicating that emodin may be a promising therapeutic option for HCC. Graphical Abstracthttps://doi.org/10.1186/s13020-025-01146-6Hepatocellular carcinomaEmodinPD-L1GSK-3β
spellingShingle Xuemei Yang
Weiguang Chen
Haitao Sun
Weicong Chen
Wei Xu
Chunyu He
Yang Liu
Ying Kuang
Yanhao Ma
Binglian Zhong
Chaojie Li
Guohuan Li
Qingfeng Du
Songqi He
Emodin promotes GSK-3β-mediated PD-L1 proteasomal degradation and enhances anti-tumor immunity in hepatocellular carcinoma
Chinese Medicine
Hepatocellular carcinoma
Emodin
PD-L1
GSK-3β
title Emodin promotes GSK-3β-mediated PD-L1 proteasomal degradation and enhances anti-tumor immunity in hepatocellular carcinoma
title_full Emodin promotes GSK-3β-mediated PD-L1 proteasomal degradation and enhances anti-tumor immunity in hepatocellular carcinoma
title_fullStr Emodin promotes GSK-3β-mediated PD-L1 proteasomal degradation and enhances anti-tumor immunity in hepatocellular carcinoma
title_full_unstemmed Emodin promotes GSK-3β-mediated PD-L1 proteasomal degradation and enhances anti-tumor immunity in hepatocellular carcinoma
title_short Emodin promotes GSK-3β-mediated PD-L1 proteasomal degradation and enhances anti-tumor immunity in hepatocellular carcinoma
title_sort emodin promotes gsk 3β mediated pd l1 proteasomal degradation and enhances anti tumor immunity in hepatocellular carcinoma
topic Hepatocellular carcinoma
Emodin
PD-L1
GSK-3β
url https://doi.org/10.1186/s13020-025-01146-6
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