Inhibitory effects of limonin on the proliferation of non-small cell lung cancer cells via targeting S-phase kinase-associated protein 2
Objective Aims to investigate the inhibitory effects of limonin on the proliferation of non-small cell lung cancer (NSCLC) cells and its molecular mechanisms. Methods To investigate the inhibitory effects of limonin on NSCLC cell activity, the MTT assay was employed. Then, flow cytometry was used to...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | zho |
| Published: |
Lanzhou University Press
2024-12-01
|
| Series: | 生物医学转化 |
| Subjects: | |
| Online Access: | http://swyxzh.ijournals.cn/swyxzh/article/html/20240412?st=article_issue |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1841543715772956672 |
|---|---|
| author | Zhu Jiaxian Zhang Mengting Li Shengqing Liu Yongqiang |
| author_facet | Zhu Jiaxian Zhang Mengting Li Shengqing Liu Yongqiang |
| author_sort | Zhu Jiaxian |
| collection | DOAJ |
| description | Objective Aims to investigate the inhibitory effects of limonin on the proliferation of non-small cell lung cancer (NSCLC) cells and its molecular mechanisms. Methods To investigate the inhibitory effects of limonin on NSCLC cell activity, the MTT assay was employed. Then, flow cytometry was used to detect the cell cycle and apoptosis, while colony formation assay assessed the low-density proliferation ability of NSCLC cells. The matrigel 3D pelletization assay was performed to evaluate stem cell properties. Further, Western blotting was used to analyze the underlying molecular mechanisms of limonin inhibiting NSCLC cell proliferation. Cell thermal shift assay (CETSA) was used to demonstrate the binding capability of limonin with target proteins, and molecular docking was employed to predict the interaction sites between limonin and target proteins. Finally, the study utilized small interfering RNA (siRNA) and S-phase kinase-associated protein 2 (Skp2) overexpressing cell lines to validate the molecular mechanisms of limonin inhibiting NSCLC cell proliferation. Results Limonin inhibited NSCLC cell proliferation in a dose-dependent manner by inducing cell cycle G0/G1 phase arrest while significantly attenuating NSCLC stem cell-like properties. Mechanistically, limonin was bound to the Skp2 protein and induced its downregulation, thus promoting the accumulation of its substrate protein p27. Skp2 knockdown by using Skp2-specific siRNA markedly enhanced the inhibitory effects of limonin on NSCLC cell activity. At the same time, Skp2 overexpression attenuated the capability of limonin to inhibit NSCLC cell activity, further validating the underlying molecular mechanisms of limonin inhibiting NSCLC cell proliferation by targeting Skp2. Conclusion Limonin inhibits NSCLC cell proliferation by targeting Skp2 protein to increase the level of the tumor suppressor protein p27, providing crucial theoretical support for the development of limonin as an antitumor drug. |
| format | Article |
| id | doaj-art-afa1c5821c204d62ad2d09af4045d77f |
| institution | Kabale University |
| issn | 2096-8965 |
| language | zho |
| publishDate | 2024-12-01 |
| publisher | Lanzhou University Press |
| record_format | Article |
| series | 生物医学转化 |
| spelling | doaj-art-afa1c5821c204d62ad2d09af4045d77f2025-01-13T07:16:35ZzhoLanzhou University Press生物医学转化2096-89652024-12-01549610510.12287/j.issn.2096-8965.20240412Inhibitory effects of limonin on the proliferation of non-small cell lung cancer cells via targeting S-phase kinase-associated protein 2Zhu Jiaxian0Zhang Mengting1Li Shengqing2Liu Yongqiang3Research Center of Chinese Herbal Resource Science and Engineering, School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine, Guangzhou 510006 , Guangdong, China ；Key Laboratory of Chinese Medicinal Resource of Lingnan (Guangzhou University of Chinese Medicine),Ministry of Education, Guangzhou 510006 , Guangdong, ChinaResearch Center of Chinese Herbal Resource Science and Engineering, School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine, Guangzhou 510006 , Guangdong, China ；Key Laboratory of Chinese Medicinal Resource of Lingnan (Guangzhou University of Chinese Medicine),Ministry of Education, Guangzhou 510006 , Guangdong, ChinaResearch Center of Chinese Herbal Resource Science and Engineering, School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine, Guangzhou 510006 , Guangdong, ChinaResearch Center of Chinese Herbal Resource Science and Engineering, School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine, Guangzhou 510006 , Guangdong, China ；Key Laboratory of Chinese Medicinal Resource of Lingnan (Guangzhou University of Chinese Medicine),Ministry of Education, Guangzhou 510006 , Guangdong, China ；Dongguan Institute of Guangzhou University of Chinese Medicine, Dongguan 523808 , Guangdong, ChinaObjective Aims to investigate the inhibitory effects of limonin on the proliferation of non-small cell lung cancer (NSCLC) cells and its molecular mechanisms. Methods To investigate the inhibitory effects of limonin on NSCLC cell activity, the MTT assay was employed. Then, flow cytometry was used to detect the cell cycle and apoptosis, while colony formation assay assessed the low-density proliferation ability of NSCLC cells. The matrigel 3D pelletization assay was performed to evaluate stem cell properties. Further, Western blotting was used to analyze the underlying molecular mechanisms of limonin inhibiting NSCLC cell proliferation. Cell thermal shift assay (CETSA) was used to demonstrate the binding capability of limonin with target proteins, and molecular docking was employed to predict the interaction sites between limonin and target proteins. Finally, the study utilized small interfering RNA (siRNA) and S-phase kinase-associated protein 2 (Skp2) overexpressing cell lines to validate the molecular mechanisms of limonin inhibiting NSCLC cell proliferation. Results Limonin inhibited NSCLC cell proliferation in a dose-dependent manner by inducing cell cycle G0/G1 phase arrest while significantly attenuating NSCLC stem cell-like properties. Mechanistically, limonin was bound to the Skp2 protein and induced its downregulation, thus promoting the accumulation of its substrate protein p27. Skp2 knockdown by using Skp2-specific siRNA markedly enhanced the inhibitory effects of limonin on NSCLC cell activity. At the same time, Skp2 overexpression attenuated the capability of limonin to inhibit NSCLC cell activity, further validating the underlying molecular mechanisms of limonin inhibiting NSCLC cell proliferation by targeting Skp2. Conclusion Limonin inhibits NSCLC cell proliferation by targeting Skp2 protein to increase the level of the tumor suppressor protein p27, providing crucial theoretical support for the development of limonin as an antitumor drug.http://swyxzh.ijournals.cn/swyxzh/article/html/20240412?st=article_issuelimoninnon-small cell lung cancers-phase kinase-associated protein 2cell proliferation |
| spellingShingle | Zhu Jiaxian Zhang Mengting Li Shengqing Liu Yongqiang Inhibitory effects of limonin on the proliferation of non-small cell lung cancer cells via targeting S-phase kinase-associated protein 2 生物医学转化 limonin non-small cell lung cancer s-phase kinase-associated protein 2 cell proliferation |
| title | Inhibitory effects of limonin on the proliferation of non-small cell lung cancer cells via targeting S-phase kinase-associated protein 2 |
| title_full | Inhibitory effects of limonin on the proliferation of non-small cell lung cancer cells via targeting S-phase kinase-associated protein 2 |
| title_fullStr | Inhibitory effects of limonin on the proliferation of non-small cell lung cancer cells via targeting S-phase kinase-associated protein 2 |
| title_full_unstemmed | Inhibitory effects of limonin on the proliferation of non-small cell lung cancer cells via targeting S-phase kinase-associated protein 2 |
| title_short | Inhibitory effects of limonin on the proliferation of non-small cell lung cancer cells via targeting S-phase kinase-associated protein 2 |
| title_sort | inhibitory effects of limonin on the proliferation of non small cell lung cancer cells via targeting s phase kinase associated protein 2 |
| topic | limonin non-small cell lung cancer s-phase kinase-associated protein 2 cell proliferation |
| url | http://swyxzh.ijournals.cn/swyxzh/article/html/20240412?st=article_issue |
| work_keys_str_mv | AT zhujiaxian inhibitoryeffectsoflimoninontheproliferationofnonsmallcelllungcancercellsviatargetingsphasekinaseassociatedprotein2 AT zhangmengting inhibitoryeffectsoflimoninontheproliferationofnonsmallcelllungcancercellsviatargetingsphasekinaseassociatedprotein2 AT lishengqing inhibitoryeffectsoflimoninontheproliferationofnonsmallcelllungcancercellsviatargetingsphasekinaseassociatedprotein2 AT liuyongqiang inhibitoryeffectsoflimoninontheproliferationofnonsmallcelllungcancercellsviatargetingsphasekinaseassociatedprotein2 |