Validation of NEDD8-conjugating enzyme UBC12 as a new therapeutic target in lung cancerResearch in context
Background: The neddylation pathway is overactivated in human cancers. Inhibition of neddylation pathway has emerged as an attractive anticancer strategy. The mechanisms underlying neddylation overactivation in cancer remain elusive. MLN4924/Pevonedistat, a first-in-class NEDD8-activating enzyme (NA...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2019-07-01
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| Series: | EBioMedicine |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2352396419303809 |
| _version_ | 1818305741357842432 |
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| author | Lihui Li Jihui Kang Wenjuan Zhang Lili Cai Shiwen Wang Yupei Liang Yanyu Jiang Xiaojun Liu Yunjing Zhang Hongfeng Ruan Guoan Chen Mingsong Wang Lijun Jia |
| author_facet | Lihui Li Jihui Kang Wenjuan Zhang Lili Cai Shiwen Wang Yupei Liang Yanyu Jiang Xiaojun Liu Yunjing Zhang Hongfeng Ruan Guoan Chen Mingsong Wang Lijun Jia |
| author_sort | Lihui Li |
| collection | DOAJ |
| description | Background: The neddylation pathway is overactivated in human cancers. Inhibition of neddylation pathway has emerged as an attractive anticancer strategy. The mechanisms underlying neddylation overactivation in cancer remain elusive. MLN4924/Pevonedistat, a first-in-class NEDD8-activating enzyme (NAE, E1) inhibitor, exerts significant anti-tumor effects, but its mutagenic resistance remains unresolved. Methods: The expression of NEDD8-conjugating enzyme UBC12/UBE2M (E2) and NEDD8 were estimated by bioinformatics analysis and western blot in human lung cancer cell lines. The malignant phenotypes of lung cancer cells were evaluated both in vitro and in vivo upon UBC12 knockdown. Cell-cycle arrest was evaluated by quantitative proteomic analysis and propidium iodide stain and fluorescence - activated cell sorting (FACS). The growth of MLN4924 - resistant H1299 cells was also evaluated upon UBC12 knockdown. Findings: The mRNA level of UBC12 in lung cancer tissues was much higher than that in normal lung tissues, increased with disease deterioration, and positively correlated with NEDD8 expression. Moreover, the overexpression of UBC12 significantly enhanced protein neddylation modification whereas the downregulation of UBC12 reduced neddylation modification of target proteins. Functionally, neddylation inactivation by UBC12 knockdown suppressed the malignant phenotypes of lung cancer cells both in vitro and in vivo. The quantitative proteomic analysis and cell cycle profiling showed that UBC12 knockdown disturbed cell cycle progression by triggering G2 phase cell-cycle arrest. Further mechanistical studies revealed that UBC12 knockdown inhibited Cullin neddylation, led to the inactivation of CRL E3 ligases and induced the accumulation of tumor-suppressive CRL substrates (p21, p27 and Wee1) to induce cell cycle arrest and suppress the malignant phenotypes of lung cancer cells. Finally, UBC12 knockdown effectively inhibited the growth of MLN4924-resistant lung cancer cells. Interpretation: These findings highlight a crucial role of UBC12 in fine-tuned regulation of neddylation activation status and validate UBC12 as an attractive alternative anticancer target against neddylation pathway. Fund: Chinese Minister of Science and Technology grant (2016YFA0501800), National Natural Science Foundation of China (Grant Nos. 81401893, 81625018, 81820108022, 81772470, 81572340 and 81602072), Innovation Program of Shanghai Municipal Education Commission (2019-01-07-00-10-E00056), Program of Shanghai Academic/Technology Research Leader (18XD1403800), National Thirteenth Five-Year Science and Technology Major Special Project for New Drug and Development (2017ZX09304001). The funders had no role in study design, data collection, data analysis, interpretation, writing of the report. Keywords: Lung cancer, UBC12, Anticancer target, Cell-cycle arrest, Overcome drug resistance |
| first_indexed | 2024-12-13T06:31:24Z |
| format | Article |
| id | doaj.art-fa545079c34e457099510b630c672094 |
| institution | Directory Open Access Journal |
| issn | 2352-3964 |
| language | English |
| last_indexed | 2024-12-13T06:31:24Z |
| publishDate | 2019-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | EBioMedicine |
| spelling | doaj.art-fa545079c34e457099510b630c6720942022-12-21T23:56:37ZengElsevierEBioMedicine2352-39642019-07-01458191Validation of NEDD8-conjugating enzyme UBC12 as a new therapeutic target in lung cancerResearch in contextLihui Li0Jihui Kang1Wenjuan Zhang2Lili Cai3Shiwen Wang4Yupei Liang5Yanyu Jiang6Xiaojun Liu7Yunjing Zhang8Hongfeng Ruan9Guoan Chen10Mingsong Wang11Lijun Jia12Cancer Institute of Traditional Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, ChinaCancer Institute of Traditional Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, ChinaCancer Institute of Traditional Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, China; Cancer Institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, ChinaCancer Institute of Traditional Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, ChinaDepartment of Laboratory Medicine, Huadong Hospital, Affiliated to Fudan University, Shanghai, ChinaCancer Institute of Traditional Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, ChinaCancer Institute of Traditional Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, ChinaCancer Institute, Fudan University Shanghai Cancer Center, Fudan University, Shanghai, ChinaCancer Institute of Traditional Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, ChinaThe First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Traditional Chinese Medicine), Hangzhou, Zhejiang, ChinaSchool of Medicine, Southern University of Science and Technology, Shenzhen, ChinaXinhua Hospital, Shanghai Jiaotong University, Shanghai, ChinaCancer Institute of Traditional Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, China; Corresponding author at: Cancer Institute, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, China.Background: The neddylation pathway is overactivated in human cancers. Inhibition of neddylation pathway has emerged as an attractive anticancer strategy. The mechanisms underlying neddylation overactivation in cancer remain elusive. MLN4924/Pevonedistat, a first-in-class NEDD8-activating enzyme (NAE, E1) inhibitor, exerts significant anti-tumor effects, but its mutagenic resistance remains unresolved. Methods: The expression of NEDD8-conjugating enzyme UBC12/UBE2M (E2) and NEDD8 were estimated by bioinformatics analysis and western blot in human lung cancer cell lines. The malignant phenotypes of lung cancer cells were evaluated both in vitro and in vivo upon UBC12 knockdown. Cell-cycle arrest was evaluated by quantitative proteomic analysis and propidium iodide stain and fluorescence - activated cell sorting (FACS). The growth of MLN4924 - resistant H1299 cells was also evaluated upon UBC12 knockdown. Findings: The mRNA level of UBC12 in lung cancer tissues was much higher than that in normal lung tissues, increased with disease deterioration, and positively correlated with NEDD8 expression. Moreover, the overexpression of UBC12 significantly enhanced protein neddylation modification whereas the downregulation of UBC12 reduced neddylation modification of target proteins. Functionally, neddylation inactivation by UBC12 knockdown suppressed the malignant phenotypes of lung cancer cells both in vitro and in vivo. The quantitative proteomic analysis and cell cycle profiling showed that UBC12 knockdown disturbed cell cycle progression by triggering G2 phase cell-cycle arrest. Further mechanistical studies revealed that UBC12 knockdown inhibited Cullin neddylation, led to the inactivation of CRL E3 ligases and induced the accumulation of tumor-suppressive CRL substrates (p21, p27 and Wee1) to induce cell cycle arrest and suppress the malignant phenotypes of lung cancer cells. Finally, UBC12 knockdown effectively inhibited the growth of MLN4924-resistant lung cancer cells. Interpretation: These findings highlight a crucial role of UBC12 in fine-tuned regulation of neddylation activation status and validate UBC12 as an attractive alternative anticancer target against neddylation pathway. Fund: Chinese Minister of Science and Technology grant (2016YFA0501800), National Natural Science Foundation of China (Grant Nos. 81401893, 81625018, 81820108022, 81772470, 81572340 and 81602072), Innovation Program of Shanghai Municipal Education Commission (2019-01-07-00-10-E00056), Program of Shanghai Academic/Technology Research Leader (18XD1403800), National Thirteenth Five-Year Science and Technology Major Special Project for New Drug and Development (2017ZX09304001). The funders had no role in study design, data collection, data analysis, interpretation, writing of the report. Keywords: Lung cancer, UBC12, Anticancer target, Cell-cycle arrest, Overcome drug resistancehttp://www.sciencedirect.com/science/article/pii/S2352396419303809 |
| spellingShingle | Lihui Li Jihui Kang Wenjuan Zhang Lili Cai Shiwen Wang Yupei Liang Yanyu Jiang Xiaojun Liu Yunjing Zhang Hongfeng Ruan Guoan Chen Mingsong Wang Lijun Jia Validation of NEDD8-conjugating enzyme UBC12 as a new therapeutic target in lung cancerResearch in context EBioMedicine |
| title | Validation of NEDD8-conjugating enzyme UBC12 as a new therapeutic target in lung cancerResearch in context |
| title_full | Validation of NEDD8-conjugating enzyme UBC12 as a new therapeutic target in lung cancerResearch in context |
| title_fullStr | Validation of NEDD8-conjugating enzyme UBC12 as a new therapeutic target in lung cancerResearch in context |
| title_full_unstemmed | Validation of NEDD8-conjugating enzyme UBC12 as a new therapeutic target in lung cancerResearch in context |
| title_short | Validation of NEDD8-conjugating enzyme UBC12 as a new therapeutic target in lung cancerResearch in context |
| title_sort | validation of nedd8 conjugating enzyme ubc12 as a new therapeutic target in lung cancerresearch in context |
| url | http://www.sciencedirect.com/science/article/pii/S2352396419303809 |
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