Mp05-12 Protein Kinase D1 Mitigates Against Etoposide Induced Dna Damage Through Up Regulation of Alpha-Catenin

You have accessJournal of UrologyProstate Cancer: Basic Research & Pathophysiology I (MP05)1 May 2024MP05-12 PROTEIN KINASE D1 MITIGATES AGAINST ETOPOSIDE INDUCED DNA DAMAGE THROUGH UP REGULATION OF ALPHA-CATENIN Sanjeev Shukla, Mohammed Al-Toubat, Samuel Serrano, Teruko Osumi, and K.C. Balaji Sanjeev ShuklaSanjeev Shukla , Mohammed Al-ToubatMohammed Al-Toubat , Samuel SerranoSamuel Serrano , Teruko OsumiTeruko Osumi , and K.C. BalajiK.C. Balaji View All Author Informationhttps://doi.org/10.1097/01.JU.0001008740.27639.cc.12AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The E-cadherin and beta-catenin interaction at the cell cells adherens junction plays a key role in cell adhesion and alteration in the expression and function of these genes is associated with disease progression in several solid tumors including prostate cancer. The membranous beta-catenin is linked to the cellular cytoskeleton through interaction with alpha-catenin at the threonine 120 (T120) residue. In this study, we demonstrate decreased beta-catenin T120 phosphorylation due to loss of PrKD1 results in uncoupling of beta- and alpha-catenin interaction. METHODS: Prostate cancer cells; LNCaP, LNCaP (ShPrKD1; silenced PrKD1), C4-2 and C4-2 PrKD1 were used for various sets of experiment to determine the role of DNA damage in PrKD1 overexpression and silencing cells. These cells were treated with varying concentrations and durations of compound-10 treatment and Etoposide (30µM), total cell lysates were prepared using RIPA Lysis and Extraction Buffer, cytosolic and nuclear fractions of C4-2 and C4-2 PrKD1 cells separated. The animal experiments were carried out following the Institutional Animal Care and Use Committee (IACUC) approval. The aim was to determine the tolerability of compound-10 by mice and generate preliminary data on efficacy of compound-10 in modulating the α-catenin and PrKD1 expressions in inhibiting tumor progression. RESULTS: In silico analysis, confirmed loss of T120 phosphorylation allows the beta- and alpha-catenin uncoupling. Forced expression of PrKD1 expression in prostate cancer cells increased alpha-catenin protein levels and was associated with resistance to etoposide induced DNA damage. Down regulation of alpha-catenin abrogates the PrKD1 protective effect of DNA damage. The in vitro results were corroborated in vivo using mouse prostate cancer xenograft model by inhibition of PrKD1 kinase activity with Compound 10, a selective PrKD inhibitor, demonstrating decreased beta-catenin T120 phosphorylation and alpha-catenin levels. CONCLUSIONS: The study for the first time provides a mechanistic basis of the protective role of PrKD1 against DNA damage. The findings could be used for further understanding of DNA damage in prostate cancer and d siRNA α-Catenin for 48 hours alone and in combinations. Source of Funding: N/A © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e47 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Sanjeev Shukla More articles by this author Mohammed Al-Toubat More articles by this author Samuel Serrano More articles by this author Teruko Osumi More articles by this author K.C. Balaji More articles by this author Expand All Advertisement PDF downloadLoading ...