PD02-01 NUAK1 AND -2 PROMOTE CONTRACTION, PROLIFERATION AND SUPPRESSION OF CELL DEATH IN HUMAN PROSTATE STROMAL CELLS: EVIDENCE FROM ISOFORM-SPECIFIC SILENCING

Abstract

You have accessJournal of UrologyCME1 Apr 2023PD02-01 NUAK1 AND -2 PROMOTE CONTRACTION, PROLIFERATION AND SUPPRESSION OF CELL DEATH IN HUMAN PROSTATE STROMAL CELLS: EVIDENCE FROM ISOFORM-SPECIFIC SILENCING Yuhan Liu, Ru Huang, Ruixiao Wang, Alexander Tamalunas, Raphaela Waidelich, Frank Strittmatter, Christian G. Stief, and Martin Hennenberg Yuhan LiuYuhan Liu More articles by this author , Ru HuangRu Huang More articles by this author , Ruixiao WangRuixiao Wang More articles by this author , Alexander TamalunasAlexander Tamalunas More articles by this author , Raphaela WaidelichRaphaela Waidelich More articles by this author , Frank StrittmatterFrank Strittmatter More articles by this author , Christian G. StiefChristian G. Stief More articles by this author , and Martin HennenbergMartin Hennenberg More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003219.01AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: In benign prostatic hyperplasia (BPH), contraction and growth in the prostate contribute to urethral obstruction and voiding symptoms. NUAK1 and -2 are serine/threonine kinases and promote myosin light chain phosphorlyation, actin organization, proliferation and suppression of cell death in non-muscle cells, which are critical for smooth muscle contraction and growth. Here, we examined effects of NUAK silencing on contraction and growth-related functions of human prostate stromal cells (WPMY-1). METHODS: Effects of NUAK1 and -2 silencing on matrix plug contraction, proliferation (EdU, Ki-67 mRNA), apoptosis and cell death (flowcytometry), viability (CCK-8) and actin organization (phalloidin) were examined in cultured WPMY-1 cells. Effects of NUAK inhibitors were examined in matrix contraction and in colony formation assays. RESULTS: Silencing of NUAK1 and -2 by transfection of WPMY-1 cells with isoform-specific siRNAs was confirmed by RT-PCR. Effects of silencing were most pronounced on proliferation and cell death. Proliferation rates were decreased by 60% and 70% by silencing of NUAK1 and NUAK2 (compared to scramble siRNA-transfected controls, 48 h after transfection), paralleled by decreases in Ki-67 mRNA by 75% and 77%. Numbers of dead cells amounted to 2.8 fold of scramble-transfected controls after silencing of NUAK1, and 4.9 fold of scramble controls after NUAK2 silencing, while no effect on apoptosis was observed. Viability was reduced by 31% and 49%, respectively, 24 h after transfection with NUAK1 and NUAK2 siRNA, compared to scramble controls. Polymerized actin was decreased by 66% and 70% by silencing of NUAK1 and -2. Time-dependent contractions in matrix contraction assays were reduced up to 45% by silencing of NUAK1, and up to 58% by silencing of NUAK2. The presumed NUAK inhibitors HTH01-015 and WZ4003 (both 10 µM) reduced contractions of WPMY-1 cells by up to 54% and 57%, and reduced the number of colonies in colony formation assays up to 78% and 64%, respectively. CONCLUSIONS: NUAK1 and -2 suppress cell death, and promote proliferation and contraction in prostate stromal cells. A role of NUAKs for BPH-relevant prostate functions driving voiding symptoms appears possible. NUAK inhibitors mimic effects of silencing on contraction, and inhibit the growth of stromal cells. Source of Funding: Deutsche Forschungsgemeinschaft (DFG), grant HE 5825/9-1; China Scholarship Council (CSC), grant 201908440478. Conflicts of Interest: Nothing to disclose © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e69 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Yuhan Liu More articles by this author Ru Huang More articles by this author Ruixiao Wang More articles by this author Alexander Tamalunas More articles by this author Raphaela Waidelich More articles by this author Frank Strittmatter More articles by this author Christian G. Stief More articles by this author Martin Hennenberg More articles by this author Expand All Advertisement PDF downloadLoading ...