Aberrant accumulation of NIK promotes tumor growth by dysregulating translation and post-translational modifications in breast cancer
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...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2023-04-01
|
| Series: | Cancer Cell International |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12935-023-02904-y |
| _version_ | 1797853577239068672 |
|---|---|
| author | Yusuke Hayashi Jun Nakayama Mizuki Yamamoto Masashi Maekawa Shinya Watanabe Shigeki Higashiyama Jun-ichiro Inoue Yusuke Yamamoto Kentaro Semba |
| author_facet | Yusuke Hayashi Jun Nakayama Mizuki Yamamoto Masashi Maekawa Shinya Watanabe Shigeki Higashiyama Jun-ichiro Inoue Yusuke Yamamoto Kentaro Semba |
| author_sort | Yusuke Hayashi |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-09T19:51:46Z |
| format | Article |
| id | doaj.art-49c64eebe1bf4dc281eb06637093e2ad |
| institution | Directory Open Access Journal |
| issn | 1475-2867 |
| language | English |
| last_indexed | 2024-04-09T19:51:46Z |
| publishDate | 2023-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Cancer Cell International |
| spelling | doaj.art-49c64eebe1bf4dc281eb06637093e2ad2023-04-03T05:41:32ZengBMCCancer Cell International1475-28672023-04-0123111710.1186/s12935-023-02904-yAberrant accumulation of NIK promotes tumor growth by dysregulating translation and post-translational modifications in breast cancerYusuke Hayashi0Jun Nakayama1Mizuki Yamamoto2Masashi Maekawa3Shinya Watanabe4Shigeki Higashiyama5Jun-ichiro Inoue6Yusuke Yamamoto7Kentaro Semba8Department of Life Science and Medical Bioscience, School of Advanced Science and Engineering, Waseda UniversityDepartment of Life Science and Medical Bioscience, School of Advanced Science and Engineering, Waseda UniversityResearch Center for Asian Infectious Diseases, The Institute of Medical Science, The University of TokyoDivision of Cell Growth and Tumor Regulation, Proteo-Science Center, Ehime UniversityTranslational Research Center, Fukushima Medical UniversityDivision of Cell Growth and Tumor Regulation, Proteo-Science Center, Ehime UniversityResearch Platform Office, The Institute of Medical Science, The University of TokyoLaboratory of Integrative Oncology, National Cancer Center Research InstituteDepartment of Life Science and Medical Bioscience, School of Advanced Science and Engineering, Waseda UniversityAbstract 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.https://doi.org/10.1186/s12935-023-02904-yNIKNon-canonical NF-κBTranslationPost-translational regulationIn vivo selectionOrthotopic xenograft |
| spellingShingle | Yusuke Hayashi Jun Nakayama Mizuki Yamamoto Masashi Maekawa Shinya Watanabe Shigeki Higashiyama Jun-ichiro Inoue Yusuke Yamamoto Kentaro Semba Aberrant accumulation of NIK promotes tumor growth by dysregulating translation and post-translational modifications in breast cancer Cancer Cell International NIK Non-canonical NF-κB Translation Post-translational regulation In vivo selection Orthotopic xenograft |
| title | Aberrant accumulation of NIK promotes tumor growth by dysregulating translation and post-translational modifications in breast cancer |
| title_full | Aberrant accumulation of NIK promotes tumor growth by dysregulating translation and post-translational modifications in breast cancer |
| title_fullStr | Aberrant accumulation of NIK promotes tumor growth by dysregulating translation and post-translational modifications in breast cancer |
| title_full_unstemmed | Aberrant accumulation of NIK promotes tumor growth by dysregulating translation and post-translational modifications in breast cancer |
| title_short | Aberrant accumulation of NIK promotes tumor growth by dysregulating translation and post-translational modifications in breast cancer |
| title_sort | aberrant accumulation of nik promotes tumor growth by dysregulating translation and post translational modifications in breast cancer |
| topic | NIK Non-canonical NF-κB Translation Post-translational regulation In vivo selection Orthotopic xenograft |
| url | https://doi.org/10.1186/s12935-023-02904-y |
| work_keys_str_mv | AT yusukehayashi aberrantaccumulationofnikpromotestumorgrowthbydysregulatingtranslationandposttranslationalmodificationsinbreastcancer AT junnakayama aberrantaccumulationofnikpromotestumorgrowthbydysregulatingtranslationandposttranslationalmodificationsinbreastcancer AT mizukiyamamoto aberrantaccumulationofnikpromotestumorgrowthbydysregulatingtranslationandposttranslationalmodificationsinbreastcancer AT masashimaekawa aberrantaccumulationofnikpromotestumorgrowthbydysregulatingtranslationandposttranslationalmodificationsinbreastcancer AT shinyawatanabe aberrantaccumulationofnikpromotestumorgrowthbydysregulatingtranslationandposttranslationalmodificationsinbreastcancer AT shigekihigashiyama aberrantaccumulationofnikpromotestumorgrowthbydysregulatingtranslationandposttranslationalmodificationsinbreastcancer AT junichiroinoue aberrantaccumulationofnikpromotestumorgrowthbydysregulatingtranslationandposttranslationalmodificationsinbreastcancer AT yusukeyamamoto aberrantaccumulationofnikpromotestumorgrowthbydysregulatingtranslationandposttranslationalmodificationsinbreastcancer AT kentarosemba aberrantaccumulationofnikpromotestumorgrowthbydysregulatingtranslationandposttranslationalmodificationsinbreastcancer |