211 ISOLATION, PROPAGATION, AND CHARACTERIZATION OF PROSTATE CANCER STEM CELLS FROM PRIMARY TUMORS

Abstract

You have accessJournal of UrologyStem Cell Research1 Apr 2012211 ISOLATION, PROPAGATION, AND CHARACTERIZATION OF PROSTATE CANCER STEM CELLS FROM PRIMARY TUMORS Rosa Park, Edison Zuniga, Rahuldev Bhalla, and Galina Botchkina Rosa ParkRosa Park Stony Brook, NY More articles by this author , Edison ZunigaEdison Zuniga Stony Brook, NY More articles by this author , Rahuldev BhallaRahuldev Bhalla Stony Brook, NY More articles by this author , and Galina BotchkinaGalina Botchkina Stony Brook, NY More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.264AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Survival for localized prostate cancer is high with surgery and radiation therapy. However, survival rates with metastatic prostate cancer remains low even with anti-androgen treatment. This limited effectiveness of standard anti-cancer therapies is attributed to the presence of prostate cancer stem cells (CSCs), which may be responsible for tumor initiation, development, invasion and response to treatment. Our goal is to identify and characterize an in vitro prostate cancer stem cell model for the evaluation of prostate CSC targeted drug effects. METHODS Samples of prostate cancer tissue were collected from 16 patients after radical prostatectomy. The harvested tissues were dissociated by enzymatic digestion in serum-free RPMI medium containg collagenases type II and type IV (Sigma-Aldrich). The dissociated cells were plated on both collagen I coated plates and nonadherent plates to grow single cell suspensions and 3D spheroids in serum free medium for propagation. A portion of these cells was injected into NOD/SCID to grow tumor xenografts. The cells were sorted and characterized under FACS and MACS analysis using markers CD133, CD44, CD166, CXCR and EpCAM. Other immunofluorescence staining and PCR arrays were conducted to further characterize cells. RESULTS 1 of 16 patient samples showed high tumorigenic capacity. The patient derived cells showed high expression of CD133, CXCR4, vimentin, and nestin. They demonstrated significantly higher tumor initiating capacity after transplantation into NOD/SCID mice with rapid tumor growth, and highest sphere-forming and clonogenic capacities compared to other cell phenotypes. Proteomics study of CSC enriched cells showed 51 up-regulated and 33 down-regulated proteins, many of which are involved in stem cell function, invasion, and metastasis, like vimentin and nestin. CONCLUSIONS Isolation of prostate cancer stem cells from primary tumors is difficult. However, we were able to demonstrate “stemness” by studying its morphology and proteonomic characteristics. Currently in vitro studies and in vivo studies of prostate CSCs using Docetaxel and SB-T-1214, developed by Stony Brook, are underway. Further efforts are needed to make patient derived stem cell lines in order to discover new CSC targeted therapies for androgen-independent prostate cancer. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e88 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information Rosa Park Stony Brook, NY More articles by this author Edison Zuniga Stony Brook, NY More articles by this author Rahuldev Bhalla Stony Brook, NY More articles by this author Galina Botchkina Stony Brook, NY More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...