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Now published in The Lancet

Validation of NEDD8-Conjugating Enzyme UBC12 As a New Therapeutic Target in Lung Cancer

29 Pages Posted: 15 May 2019

See all articles by Lihui Li

Lihui Li

Shanghai University of Traditional Chinese Medicine - Cancer Institute

Jihui Kang

Shanghai University of Traditional Chinese Medicine - Cancer Institute

Wenjuan Zhang

Shanghai University of Traditional Chinese Medicine - Cancer Institute; Fudan University - Shanghai Cancer Center (FUSCC)

Lili Cai

Shanghai University of Traditional Chinese Medicine - Cancer Institute

Shiwen Wang

Fudan University - Huadong Hospital

Yupei Liang

Shanghai University of Traditional Chinese Medicine - Cancer Institute

Yanyu Jiang

Shanghai University of Traditional Chinese Medicine - Cancer Institute

Xiaojun Liu

Fudan University - Shanghai Cancer Center (FUSCC)

Yunjing Zhang

Shanghai University of Traditional Chinese Medicine - Cancer Institute

Hongfeng Ruan

Zhejiang Chinese Medical University - The First Affiliated Hospital

Guoan Chen

Southern University of Science and Technology - School of Medicine

Mingsong Wang

Shanghai Jiao Tong University (SJTU) - Xinhua Hospital

Lijun Jia

Shanghai Jiao Tong University (SJTU) - Department of Oncology; Shanghai University of Traditional Chinese Medicine - Cancer Institute

More...

Abstract

Backgrounds: 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 was estimated by bioinformatics analysis and western blotting 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.

Results: 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.

Conclusions: 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.

Funding Statement: This work was supported by the 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).

Declaration of Interests: The authors declare that they have no competing interests.

Ethics Approval Statement: The authors state: "Not applicable."

Keywords: Lung cancer, UBC12, anticancer target, cell-cycle arrest, overcome drug resistance

Suggested Citation

Li, Lihui and Kang, Jihui and Zhang, Wenjuan and Cai, Lili and Wang, Shiwen and Liang, Yupei and Jiang, Yanyu and Liu, Xiaojun and Zhang, Yunjing and Ruan, Hongfeng and Chen, Guoan and Wang, Mingsong and Jia, Lijun, Validation of NEDD8-Conjugating Enzyme UBC12 As a New Therapeutic Target in Lung Cancer (May 13, 2019). Available at SSRN: https://ssrn.com/abstract=3387508 or http://dx.doi.org/10.2139/ssrn.3387508

Lihui Li

Shanghai University of Traditional Chinese Medicine - Cancer Institute

Shanghai, 200032
China

Jihui Kang

Shanghai University of Traditional Chinese Medicine - Cancer Institute

Shanghai, 200032
China

Wenjuan Zhang

Shanghai University of Traditional Chinese Medicine - Cancer Institute

Shanghai, 200032
China

Fudan University - Shanghai Cancer Center (FUSCC)

1207 Rm., 2# Bldg., 270 Dong An Rd.
Shanghai, 200032
China

Lili Cai

Shanghai University of Traditional Chinese Medicine - Cancer Institute

Shanghai, 200032
China

Shiwen Wang

Fudan University - Huadong Hospital

Shanghai
China

Yupei Liang

Shanghai University of Traditional Chinese Medicine - Cancer Institute

Shanghai, 200032
China

Yanyu Jiang

Shanghai University of Traditional Chinese Medicine - Cancer Institute

Shanghai, 200032
China

Xiaojun Liu

Fudan University - Shanghai Cancer Center (FUSCC)

1207 Rm., 2# Bldg., 270 Dong An Rd.
Shanghai, 200032
China

Yunjing Zhang

Shanghai University of Traditional Chinese Medicine - Cancer Institute

Shanghai, 200032
China

Hongfeng Ruan

Zhejiang Chinese Medical University - The First Affiliated Hospital

Hangzhou, Zhejiang
China

Guoan Chen

Southern University of Science and Technology - School of Medicine ( email )

No 1088, xueyuan Rd.
Xili, Nanshan District
Shenzhen, Guangdong 518055
China

Mingsong Wang

Shanghai Jiao Tong University (SJTU) - Xinhua Hospital

Shanghai
China

Lijun Jia (Contact Author)

Shanghai Jiao Tong University (SJTU) - Department of Oncology

Shanghai
China

Shanghai University of Traditional Chinese Medicine - Cancer Institute ( email )

Shanghai, 200032
China

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