Puerarin modulation of CENPA affects downstream PLK1 and CCNB1 expression to inhibit bladder cancer cell proliferation  

作　　者：Hao Xu Wen Gao Deng Pan Yu-Yang Ma Ruo-Ran Zhang Yong-Li Cao Yu-Chuan Zhou Ming-Yu Xu Pei-Yong Zhang Kun Pang 

机构地区：[1]Department of Urology,Xuzhou Central Hospital,Xuzhou Clinical School of Xuzhou Medical College,Xuzhou 221009,China [2]Graduate School,Bengbu Medical University,Bengbu 233040,China [3]Department of Cardiology,The Fourth People’s Hospital of Jinan,Jinan 250031,China [4]Graduate School,Jiangsu University,Zhenjiang 212013,China [5]Department of Urology,The Peixian People’s Hospital,Xuzhou 221699,China [6]Department of Clinical Laboratory,The First Affiliated Hospital of Harbin Medical University,Harbin 150006,China

出　　处：《Traditional Medicine Research》2024年第9期32-41,共10页TMR传统医学研究

基　　金：supported by National Natural Science Fund Item Number(82004110);Xuzhou Science and Technology Plan Project(KC22096);China Postdoctoral Science Foundation(2022M722674);Xuzhou Medical Reserve Talents Project(XWRCHT20220009);the Xuzhou Clinical Medicine Expert Team Project(2018TD004);Peixian Science and Technology Program(P202410)。

摘　　要：Background:The treatment alternatives for bladder cancer(BLCA),the 10th most prevalent cancer in the world,need to be further investigated,and many active substances like Puerarin in herbal medicine were found to be effective in treating BLCA.The purpose of this study was to investigate the potential treating mechanisms of Puerarin on BLCA.Methods:The cell counting kit 8 assay and flow cytometry were performed to confirm Puerarin’s ability to suppress BLCA.The differentially expressed proteins(DEPs)were obtained by Tandem Mass Tags technology and functional enrichment analysis performed by R studio.The most enriched pathways were selected for study and the DEPs were screened out.Protein-protein interaction network maps were created using String and Cytoscape and key proteins,which will be analyzed for survival,expression,and upstream transcription factor prediction,were screened out using the cytoHubba plugin.CHEA3 was used to obtain upstream transcription factor validated by molecular docking and western blotting experiments.Results:Cell counting kit 8 showed that Puerarin inhibited BLCA cells,with 50%inhibitory concentration of 218μmol/L in T24 and 198μmol/L in 5637.Flow cytometry reveals that Puerarin blocks T24 and 5637 cells in G1 phase.1,385 DEPs were obtained and the enrichment analysis revealed that cell cycle and DNA replication were the two main areas in which DEPs were enriched.Cyclin-B-cyclin dependent kinase 1(CDK1),cyclin B1(CCNB1),and polo-like kinase 1(PLK1)were identified as key proteins,and their upstream transcription factor was predicted to be centromere protein A(CENPA).Puerarin’s binding energy to CENPA was determined by molecular docking to be−6.3 kcal/mol,indicating a strong binding interaction.Western blot showed that Puerarin significantly reduced the expression of CENPA.Conclusion:We hypothesize that Puerarin may inhibit the proliferation of bladder cancer cells by inhibiting CENPA expression to regulate PLK1 and CCNB1 expression,thereby affecting cell cycle.

关 键 词：PUERARIN BLCA Tandem Mass Tags cell cycle 

分 类 号：R737.14[医药卫生—肿瘤]