EZH2 mediates mechanisms of AD20 on its inhibitory effects in PC3 cell proliferation and migration

摘要

OBJECTIVE Prostate cancer is the most commonly diagnosed malignancy and the sixth leading cause of cancer death in men.Aberrant upregulation of enhancer of Zeste homolog2(EZH2)occur frequently in human cancers,including prostate adenocarcinoma,it′s a bona fide oncogene,yet clinical benefits of EZH2 inhibitor(EZH2i)remain unsatis⁃factory and limited to certain hematological malignancies.AD20 is a natural product which is extracted from creosote bush.Our previous study has demonstrated that AD20 could inhibit the growth and metastasis in prostate cancer by inhibiting NRP1,however,the mechanisms still need to be clarified.METHODS PC3 cell were cultured in DMEM/F12 supplemented with 10%fetal bovine serum.The proliferation and migration were measured by MTS,wound healing assay and tran⁃swell assay.The expression of the protein was observed by Western blot.The mRNA levels were detected by real-time PCR.LC-MS/MS-based proteomic assay,CoIP-MS and network pharmacology are used to construct the PPI network and identify related proteins.RESULTS AD20(10-20μmol·L^-1)can regulate EZH2 expression from transcription,trans⁃lation,degradation levels.AD20 downregulates EZH2-H3K27me3 signaling dramatically and activates many down⁃stream tumor suppressor genes,such as CDH1,GATA6,CDKN2A.In vitro,AD20(10-40μmol·L^-1)suppresses PC3 cell proliferation and migration in a dose-dependent manner.And interestingly,the combination with AD20 significantly improved the efficacy of EZH2 inhibitors(GSK126 and EPZ-6438)in PC3 cell line.CONCLUSION AD20 can suppress PC3 cell proliferation by targeting EZH2-H3K27me3-p16INK4a signaling and inhibit PC3 cell migration by inhibiting NRP1.