类泛素化neddylation修饰调控乳腺癌中核纤层蛋白lamin B1的表达

doi:10.19401/j.cnki.1007-3639.2022.06.008

摘要/Abstract

摘要：

背景与目的：类泛素化neddylation修饰在乳腺癌中的作用鲜有报道。该研究的前期研究发现,抑制neddylation修饰可诱导乳腺癌细胞发生衰老,但具体机制尚未完全阐明。研究报道核纤层蛋白lamin B1缺失可导致细胞衰老。本研究旨在探讨在乳腺癌中,neddylation修饰与lamin B1是否存在相关性及其可能的机制。方法：利用113例乳腺癌患者肿瘤组织制成的组织芯片对neddylation修饰过程中关键蛋白神经前体细胞表达发育下调蛋白8（neural precursor cell expressed developmentally downregulated protein 8,NEDD8）、NEDD8 活化酶E1（NEDD8-activating enzyme 1,NAE1）及核纤层蛋白lamin B1进行免疫组织化学染色。采用Spearman rank分析NEDD8和NAE1与laminB1表达的相关性。采用CRISPR/Cas9技术敲低NEDD8的表达以阻断neddylation修饰,采用蛋白质印迹法（Western blot）检测neddylation修饰对Lamin B1表达的调控作用及其机制。结果：Lamin B1表达与NEDD8（r=0.817,P<0.000 1）和NAE1（r=0.406,P<0.000 1）表达呈显著正相关。敲低NEDD8以阻断neddylation修饰可显著抑制Lamin B1的表达。沉默P53表达可部分逆转阻断neddylation修饰对lamin B1表达的抑制。结论：Neddylation修饰与lamin B1表达呈正相关,靶向neddylation修饰可能通过P53依赖的方式抑制lamin B1的表达。这为乳腺癌的治疗提供了新的靶点和方向。

关键词: 类泛素化, Neddylation, Lamin B1, 乳腺癌

Abstract:

Background and purpose: The role of neddylation modification in breast cancer has rarely been reported. Previous study found that neddylation modification blocking induced cellular senescence in breast cancer, however, the potential mechanism has not yet been fully elucidated. It is reported that lamin B1 deletion leads to cellular senescence. The purpose of this study was to investigate the correlation between neddylation modification and lamin B1 expression in breast cancer and its possible mechanism. Methods: Neural precursor cell expressed developmentally downregulated protein 8 (NEDD8) and NEDD8-activating enzyme 1 (NAE1), two key proteins of neddylation modification process, in addition to lamin B1 were immunohistochemically stained in tissue microarray from 113 breast cancer patients. The correlation of NEDD8 and NAE1 with lamin B1 was analyzed using Spearman’s rank analysis. The expression of NEDD8 was knocked down to block neddylation modification by CRISPR/Cas9 technique. Western blot assay was used to detect the regulation of lamin B1 by neddylation modification and its mechanisms. Results: Lamin B1 expression was positively correlated with NEDD8 (r=0.817, P<0.000 1) and NAE1 (r=0.406, P<0.000 1) expressions. Blocking neddylation modification by knocking down NEDD8 significantly inhibited the expression of lamin B1. Silencing of p53 partially reversed the inhibition of lamin B1 by neddylation modification blocking. Conclusion: Neddylation modification was positively correlated with lamin B1 expression. Targeting neddylation modification inhibited the expression of lamin B1 in a p53-dependent manner. This study provides new target and clue for the treatment of breast cancer.

Key words: Ubiquitin-like, Neddylation, Lamin B1, Breast cancer

中图分类号:

R737.9

贾晓青, 孙晓寅, 蒋蓓琦, 梅章懿, 傅韵, 莊志刚. 类泛素化neddylation修饰调控乳腺癌中核纤层蛋白lamin B1的表达[J]. 中国癌症杂志, 2022, 32(6): 535-541.

JIA Xiaoqing, SUN Xiaoyin, JIANG Beiqi, MEI Zhangyi, FU Yun, ZHUANG Zhigang. Neddylation modification regulates lamin B1 expression in breast cancer[J]. China Oncology, 2022, 32(6): 535-541.

图/表 5

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