过度激活的 neddylation 通路作为肺癌的治疗靶点。
Overactivated neddylation pathway as a therapeutic target in lung cancer.
机构信息
Affiliations of authors: Cancer Institute, Fudan University Shanghai Cancer Center (LL, GY, PC, DW, CL, WY, YW, LJ), Department of Oncology, Shanghai Medical College (LL, GY, PC, DW, CL, WY, YW, LJ), Department of Immunology, School of Basic Medical Sciences (LL, GY, CL, YW, YC, LJ), Clinical Statistics Center, Department of Radiation Oncology, Fudan University Shanghai Cancer Center (JZ, LX, HJ), and Liver Cancer Institute, Zhongshan Hospital (JS, QG), Fudan University, Shanghai, 200032, China; Department of Thoracic Cardiovascular Surgery, Xinhua Hospital of Shanghai Jiaotong University School of Medicine, Shanghai, China (MW, FH, JM); College of Basic Medical Sciences, Zhengzhou University, Zhengzhou, China (PC, ZD); Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, China (HL, PW); College of Pharmacy, Seoul National University, Seoul, Korea (LSJ); College of Pharmacy, Ewha Womans University, Seoul, Korea (LSJ, HWL, JY); AntiCancer Biotech Beijing Co. Ltd., Beijing, China (HQ, MY); Division of Radiation and Cancer Biology, Department of Radiation Oncology, University of Michigan, Ann Arbor, MI (YS); Department of Surgery, University of California, San Diego, CA (RMH); AntiCancer, Inc., San Diego, CA (RMH, MY).
出版信息
J Natl Cancer Inst. 2014 May 22;106(6):dju083. doi: 10.1093/jnci/dju083. Print 2014 Jun.
BACKGROUND
A number of oncoproteins and tumor suppressors are known to be neddylated, but whether the neddylation pathway is entirely activated in human cancer remains unexplored.
METHODS
NEDD8-activating enzyme (NAE) (E1) and NEDD8-conjugating enzyme (E2) expression and global-protein neddylation were examined by immunohistochemistry, immunoblotting, and real-time polymerase chain reaction analysis. Cell proliferation, clonogenic survival, migration, and motility in vitro, as well as tumor formation and metastasis in vivo, were determined upon neddylation inhibition by MLN4924, an investigational NEDD8-activating enzyme inhibitor. Survival was analyzed with Kaplan-Meier methods and compared by the log-rank test. All statistical tests were two-sided.
RESULTS
The entire neddylation pathway, including NEDD8-activating enzyme E1, NEDD8-conjugating enzyme E2, and global-protein neddylation, is overactivated in both lung adenocarcinoma and squamous-cell carcinoma. Compared with lung adenocarcinoma patients with low expression, those with high expression had worse overall survival (NEDD8-activating enzyme E1 subunit 1 [NAE1]: hazard ratio [HR] = 2.07, 95% confidence interval [CI] = 0.95 to 4.52, P = .07; ubiquitin-conjugating enzyme E2M (UBC12): HR = 13.26, 95% CI = 1.77 to 99.35, P = .01; global protein neddylation: HR = 3.74, 95% CI = 1.65 to 8.47, P = .002). Moreover, inhibition of neddylation by the NAE inhibitor MLN4924 statistically significantly suppressed proliferation, survival, migration, and motility of lung cancer cells in vitro and tumor formation and metastasis in vivo. At the molecular level, MLN4924 inactivated Cullin-RING E3 ligases, led to accumulation of tumor-suppressive Cullin-RING E3 ligase substrates and induced phorbol-12-myristate-13-acetate-induced protein 1 (NOXA)-dependent apoptosis or cellular senescence.
CONCLUSIONS
Our study highlights the overactivated neddylation pathway in lung cancer development and as a promising therapeutic target.
背景
已知许多癌蛋白和肿瘤抑制因子都被 neddylation，但是否在人类癌症中完全激活了 neddylation 途径仍未被探索。
方法
通过免疫组织化学、免疫印迹和实时聚合酶链反应分析，检测 NEDD8 激活酶（E1）和 NEDD8 连接酶（E2）的表达和全蛋白 neddylation。通过 MLN4924 抑制 neddylation（一种研究性的 NEDD8 激活酶抑制剂），检测体外细胞增殖、集落形成存活、迁移和运动，以及体内肿瘤形成和转移。采用 Kaplan-Meier 方法分析生存情况，并通过对数秩检验进行比较。所有统计检验均为双侧检验。
结果
在肺腺癌和鳞状细胞癌中，整个 neddylation 途径，包括 NEDD8 激活酶 E1、NEDD8 连接酶 E2 和全蛋白 neddylation，均过度激活。与低表达的肺腺癌患者相比，高表达的患者总生存期更差（NEDD8 激活酶 E1 亚基 1 [NAE1]：风险比 [HR] = 2.07，95%置信区间 [CI] = 0.95 至 4.52，P =.07；泛素连接酶 E2M（UBC12）：HR = 13.26，95%CI = 1.77 至 99.35，P =.01；全蛋白 neddylation：HR = 3.74，95%CI = 1.65 至 8.47，P =.002）。此外，NAE 抑制剂 MLN4924 抑制 neddylation 可显著抑制肺癌细胞的体外增殖、存活、迁移和运动，以及体内肿瘤形成和转移。在分子水平上，MLN4924 使 Cullin-RING E3 连接酶失活，导致肿瘤抑制性 Cullin-RING E3 连接酶底物的积累，并诱导佛波醇 12-肉豆蔻酸 13-乙酸酯诱导蛋白 1（NOXA）依赖性细胞凋亡或细胞衰老。
结论
我们的研究强调了 neddylation 途径在肺癌发生发展中的过度激活，并作为一个有前途的治疗靶点。
Zotero 插件