NIK的异常积累通过失调乳腺癌中的翻译和翻译后修饰来促进肿瘤生长。

Aberrant accumulation of NIK promotes tumor growth by dysregulating translation and post-translational modifications in breast cancer.

作者信息

Hayashi Yusuke, Nakayama Jun, Yamamoto Mizuki, Maekawa Masashi, Watanabe Shinya, Higashiyama Shigeki, Inoue Jun-Ichiro, Yamamoto Yusuke, Semba Kentaro

机构信息

Department of Life Science and Medical Bioscience, School of Advanced Science and Engineering, Waseda University, TWIns, 2-2 Wakamatsu-Cho, Shinjuku-Ku, Tokyo, 162-8480, Japan.

Laboratory of Integrative Oncology, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-Ku, Tokyo, 104-0045, Japan.

出版信息

Cancer Cell Int. 2023 Apr 1;23(1):57. doi: 10.1186/s12935-023-02904-y.

DOI:10.1186/s12935-023-02904-y

PMID:37005661

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10067241/

Abstract

BACKGROUND

In vivo investigations with cancer cells have powerful tools to discover cancer progression mechanisms and preclinical candidate drugs. Among these in vivo experimental models, the establishment of highly malignancy cell lines with xenograft has been frequently used. However, few previous researches targeted malignancy-related genes whose protein levels translationally changed. Therefore, this study aimed to identify malignancy-related genes which contributed to cancer progression and changed at the protein level in the in vivo selected cancer cell lines.

METHODS

We established the high malignancy breast cancer cell line (LM05) by orthotopic xenograft as an in vivo selection method. To explore the altered genes by translational or post-translational regulation, we analyzed the protein production by western blotting in the highly malignant breast cancer cell line. Functional analyses of the altered genes were performed by in vitro and in vivo experiments. To reveal the molecular mechanisms of the regulation with protein level, we evaluated post-translational modification by immunoprecipitation. In addition, we evaluated translational production by click reaction-based purification of nascent protein.

RESULTS

As a result, NF-κB inducing kinase (NIK) increased at the protein level and promoted the nuclear localization of NF-κB2 (p52) and RelB in the highly malignant breast cancer cell line. The functional analyses indicated the NIK upregulation contributed to tumor malignancy via cancer-associated fibroblasts (CAFs) attraction and partially anti-apoptotic activities. Additionally, the immunoprecipitation experiment revealed that the ubiquitination of NIK decreased in LM05 cells. The decline in NIK ubiquitination was attributed to the translational downregulation of cIAP1.

CONCLUSIONS

Our study identified a dysregulated mechanism of NIK production by the suppression of NIK post-modification and cIAP1 translation. The aberrant NIK accumulation promoted tumor growth in the highly malignant breast cancer cell line.

摘要

背景

利用癌细胞进行体内研究是发现癌症进展机制和临床前候选药物的有力工具。在这些体内实验模型中，建立具有异种移植的高恶性细胞系被广泛应用。然而，以往很少有研究针对蛋白质水平发生翻译变化的恶性相关基因。因此，本研究旨在鉴定在体内选择的癌细胞系中有助于癌症进展且在蛋白质水平发生变化的恶性相关基因。

方法

我们通过原位异种移植建立了高恶性乳腺癌细胞系（LM05）作为体内选择方法。为了探索通过翻译或翻译后调控改变的基因，我们在高恶性乳腺癌细胞系中通过蛋白质免疫印迹分析蛋白质产生情况。通过体外和体内实验对改变的基因进行功能分析。为了揭示蛋白质水平调控的分子机制，我们通过免疫沉淀评估翻译后修饰。此外，我们通过基于点击反应的新生蛋白质纯化评估翻译产生情况。

结果

结果显示，在高恶性乳腺癌细胞系中，核因子κB诱导激酶（NIK）在蛋白质水平升高，并促进了NF-κB2（p52）和RelB的核定位。功能分析表明，NIK上调通过吸引癌症相关成纤维细胞（CAF）和部分抗凋亡活性促进肿瘤恶性程度。此外，免疫沉淀实验表明，LM05细胞中NIK的泛素化减少。NIK泛素化的下降归因于细胞凋亡抑制蛋白1（cIAP1）的翻译下调。