RNF182 induces p65 ubiquitination to affect PDL1 transcription and suppress immune evasion in lung adenocarcinoma
Abstract Background The RING finger (RNF) proteins are a large group of ubiquitin ligases whose aberrant expression is often associated with disease progression. This study examines the function of RNF protein 182 (RNF182) in lung adenocarcinoma (LUAD) cells and its impact on p65 and programmed deat...
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Wiley
2023-05-01
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| Series: | Immunity, Inflammation and Disease |
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| Online Access: | https://doi.org/10.1002/iid3.864 |
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| author | Xingdu Zeng Xiaoyuan Tang Xingxiang Chen Huilan Wen |
| author_facet | Xingdu Zeng Xiaoyuan Tang Xingxiang Chen Huilan Wen |
| author_sort | Xingdu Zeng |
| collection | DOAJ |
| description | Abstract Background The RING finger (RNF) proteins are a large group of ubiquitin ligases whose aberrant expression is often associated with disease progression. This study examines the function of RNF protein 182 (RNF182) in lung adenocarcinoma (LUAD) cells and its impact on p65 and programmed death ligand 1 (PDL1) regulation. Methods Expression of RNF182, p65, and PDL1 in LUAD tissues and cells was measured using immunohistochemistry, reverse transcription quantitative polymerase chain reaction (RT‐qPCR), and/or western blot (WB) assays. LUAD cells were induced to overexpress RNF182 and p65, followed by cell counting kit‐8, colony formation, Transwell, and flow cytometry assays to evaluate the cells’ malignant phenotype. Coimmunoprecipitation and WB assays were used to verify RNF182's effect on p65 ubiquitination. Chromatin immunoprecipitation‐qPCR and luciferase assays were used to analyze p65's transcriptional regulation of PDL1. Coculture of LUAD with CD8+ cytotoxic T cells was performed to detect lactate dehydrogenase release and interferon‐γ and interleukin‐2 concentrations. LUAD cells were implanted in mice to analyze tumorigenicity. Results RNF182 was poorly expressed, while p65 and PDL1 were highly expressed in LUAD tissues and cells. RNF182 overexpression suppressed the malignant properties of LUAD cells, and it promoted p65 ubiquitination and protein degradation. p65 activated PDL1 transcription. Overexpression of RNF182 suppressed the PDL1 expression, increased the cytotoxicity in LUAD cells cocultured with CD8+ T cells, and suppressed the tumorigenesis of cancer cells in vivo. However, these tumor‐suppressive effects of RNF182 on LUAD cells were blocked by p65 restoration. Conclusion This research demonstrates that RNF182 induces p65 ubiquitination to suppress PDL1 transcription and immunosuppression in LUAD. |
| format | Article |
| id | doaj-art-1f16cd13364d427c845abd50fbeacf6b |
| institution | OA Journals |
| issn | 2050-4527 |
| language | English |
| publishDate | 2023-05-01 |
| publisher | Wiley |
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| series | Immunity, Inflammation and Disease |
| spelling | doaj-art-1f16cd13364d427c845abd50fbeacf6b2025-08-20T02:15:41ZengWileyImmunity, Inflammation and Disease2050-45272023-05-01115n/an/a10.1002/iid3.864RNF182 induces p65 ubiquitination to affect PDL1 transcription and suppress immune evasion in lung adenocarcinomaXingdu Zeng0Xiaoyuan Tang1Xingxiang Chen2Huilan Wen3Department of Respiratory Medicine The First Affiliated Hospital of Gannan Medical University Ganzhou Jiangxi People's Republic of ChinaDepartment of Respiratory Medicine The First Affiliated Hospital of Gannan Medical University Ganzhou Jiangxi People's Republic of ChinaDepartment of Respiratory Medicine The First Affiliated Hospital of Gannan Medical University Ganzhou Jiangxi People's Republic of ChinaDepartment of Respiratory Medicine The First Affiliated Hospital of Gannan Medical University Ganzhou Jiangxi People's Republic of ChinaAbstract Background The RING finger (RNF) proteins are a large group of ubiquitin ligases whose aberrant expression is often associated with disease progression. This study examines the function of RNF protein 182 (RNF182) in lung adenocarcinoma (LUAD) cells and its impact on p65 and programmed death ligand 1 (PDL1) regulation. Methods Expression of RNF182, p65, and PDL1 in LUAD tissues and cells was measured using immunohistochemistry, reverse transcription quantitative polymerase chain reaction (RT‐qPCR), and/or western blot (WB) assays. LUAD cells were induced to overexpress RNF182 and p65, followed by cell counting kit‐8, colony formation, Transwell, and flow cytometry assays to evaluate the cells’ malignant phenotype. Coimmunoprecipitation and WB assays were used to verify RNF182's effect on p65 ubiquitination. Chromatin immunoprecipitation‐qPCR and luciferase assays were used to analyze p65's transcriptional regulation of PDL1. Coculture of LUAD with CD8+ cytotoxic T cells was performed to detect lactate dehydrogenase release and interferon‐γ and interleukin‐2 concentrations. LUAD cells were implanted in mice to analyze tumorigenicity. Results RNF182 was poorly expressed, while p65 and PDL1 were highly expressed in LUAD tissues and cells. RNF182 overexpression suppressed the malignant properties of LUAD cells, and it promoted p65 ubiquitination and protein degradation. p65 activated PDL1 transcription. Overexpression of RNF182 suppressed the PDL1 expression, increased the cytotoxicity in LUAD cells cocultured with CD8+ T cells, and suppressed the tumorigenesis of cancer cells in vivo. However, these tumor‐suppressive effects of RNF182 on LUAD cells were blocked by p65 restoration. Conclusion This research demonstrates that RNF182 induces p65 ubiquitination to suppress PDL1 transcription and immunosuppression in LUAD.https://doi.org/10.1002/iid3.864E3 ubiquitin ligaseimmune evasionNF‐kB p65PDL1RNF182 |
| spellingShingle | Xingdu Zeng Xiaoyuan Tang Xingxiang Chen Huilan Wen RNF182 induces p65 ubiquitination to affect PDL1 transcription and suppress immune evasion in lung adenocarcinoma Immunity, Inflammation and Disease E3 ubiquitin ligase immune evasion NF‐kB p65 PDL1 RNF182 |
| title | RNF182 induces p65 ubiquitination to affect PDL1 transcription and suppress immune evasion in lung adenocarcinoma |
| title_full | RNF182 induces p65 ubiquitination to affect PDL1 transcription and suppress immune evasion in lung adenocarcinoma |
| title_fullStr | RNF182 induces p65 ubiquitination to affect PDL1 transcription and suppress immune evasion in lung adenocarcinoma |
| title_full_unstemmed | RNF182 induces p65 ubiquitination to affect PDL1 transcription and suppress immune evasion in lung adenocarcinoma |
| title_short | RNF182 induces p65 ubiquitination to affect PDL1 transcription and suppress immune evasion in lung adenocarcinoma |
| title_sort | rnf182 induces p65 ubiquitination to affect pdl1 transcription and suppress immune evasion in lung adenocarcinoma |
| topic | E3 ubiquitin ligase immune evasion NF‐kB p65 PDL1 RNF182 |
| url | https://doi.org/10.1002/iid3.864 |
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