文献类型：期刊文献

英文题名：A protein-fragment complementation assay reveals that celastrol and gambogic acid suppress ERα mutants in breast cancer

作者：Liu, Xi[1];Hu, Qian[1];Wang, Wanyan[1];Ma, Hui[1];Pu, Jiaqian[1];Cui, Jiayan[1];Gong, Ting[1];Wu, Yu[1];Lu, Weiqiang[2,3];Huang, Jin[1]

机构：[1]East China Univ Sci & Technol, Shanghai Key Lab New Drug Design, Sch Pharm, Shanghai 200237, Peoples R China;[2]East China Normal Univ, Shanghai Key Lab Regulatory Biol, Inst Biomed Sci, Shanghai 200241, Peoples R China;[3]East China Normal Univ, Sch Life Sci, Shanghai 200241, Peoples R China

年份：2021

卷号：188

外文期刊名：BIOCHEMICAL PHARMACOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000656195100002)】；

基金：This work was supported by the National Natural Science Foundation of China (81973362, 81972828, 81773775) and Shanghai Committee of Science and Technology (19ZR1473500, 18431900500) . We thank the Open Research Project of Key Laboratory of HighIncidenceTumor Prevention & Treatment (Guangxi Medical University) , Ministry of Education and the Research Center of Analysis and Test of East China University of Science and Technology.

语种：英文

外文关键词：Estrogen receptor alpha; Breast cancer; GLPCA; Celastrol; Gambogic acid

摘要：Somatic gain-of-function mutations within estrogen receptor alpha (ER alpha) are highly associated with hormone therapy resistance in breast cancer. However, current understanding of abnormal activity of ER alpha mutants and their relevant targeted intervention is still very limited. Herein, we developed a new, real-time, and reliably Gaussia luciferase-based protein-fragment complementation assay (GLPCA) for evaluating ER alpha mutants activities. We found that, compared with ER WT, ER alpha mutants (Y537S/N and D538G) exhibit high ligand-independent activity, suggesting the gain-of-function phenotype of these ER alpha mutants. Notably, Y537S, the most common ER alpha mutant type, has the highest intrinsic activation. We then collected and screened a natural product library for potential ER alpha antagonists via GLPCA and identified celastrol and gambogic acid as new antagonists of the ER alpha Y537S mutant. Moreover, interactions between these two compounds and the ER alpha Y537S mutant were confirmed by molecular docking and cellular thermal shift assay. Importantly, we further demonstrated that celastrol and gambogic acid exhibit synergistic antiproliferative and pro-apoptotic effects when combined with an approved CDK4/6 inhibitor abemaciclib in breast cancer cells expressing ER alpha Y537S. In summary, GLPCA provides a powerful platform for exploring innovative functional biology and drug discovery of antagonists targeting ER alpha mutants.