Document Detail


Characterization of benign and malignant prostate epithelial Hoechst 33342 side populations.
MedLine Citation:
PMID:  17639507     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: The prostate epithelial stem cell has been proposed as the primary origin of neoplastic change in prostate cancer. However, the isolation and characterization of unexpanded prostate epithelial stem cells have proven problematic.
METHODS: A prostate epithelial side population (SP) has been isolated utilizing a modified Hoechst 33342 dye efflux assay from both benign and malignant prostate tissue. CD45(-ve), integrin alpha2(+ve) Hoechst 33342 SP and NSP cells were isolated by FACS, immunophenotyped and functionally characterized in 3D culture.
RESULTS: FACS analysis revealed a verapamil sensitive SP accounting for 0.93 +/- 0.12% and 0.57 +/- 0.11% of the total epithelial population from both benign and malignant prostates. The benign SP phenotype revealed a heterogeneous cell population consisting predominantly of small basal cells containing minimal cytoplasm. Conversely, the malignant SP was of undetermined acinar origin and with a complete loss of expression of the CDK2 inhibitor p21(WAF1/Cip1). In vitro androgen-enhanced 3D culture of the benign and malignant SP cells led to the production of spheroids which had acinus like morphology and expressed primitive and basal cell markers. Incorporation of the CD133 marker isolated a further SP sub-fraction accounting for 0.037 +/- 0.01% of epithelial cells.
CONCLUSIONS: Our observations are consistent with the Hoechst 33342 dye efflux assay isolating a stem cell enriched population which can be further sub-fractionated by CD133 selection. Moreover, the loss of the CDK inhibitor in malignancy is consistent with the hypothesis that neoplastic change originates in the stem cell compartment.
Authors:
Mick D Brown; Paul E Gilmore; Claire A Hart; Joanne D Samuel; Vijay A C Ramani; Nicholas J George; Noel W Clarke
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Prostate     Volume:  67     ISSN:  0270-4137     ISO Abbreviation:  Prostate     Publication Date:  2007 Sep 
Date Detail:
Created Date:  2007-08-02     Completed Date:  2007-10-23     Revised Date:  2014-02-19    
Medline Journal Info:
Nlm Unique ID:  8101368     Medline TA:  Prostate     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1384-96     Citation Subset:  IM    
Copyright Information:
2007 Wiley-Liss, Inc
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MeSH Terms
Descriptor/Qualifier:
Adult Stem Cells / cytology*,  metabolism,  pathology
Antigens, CD / biosynthesis
Antigens, CD45 / biosynthesis
Benzimidazoles / chemistry*
Cell Fractionation / methods
Cell Growth Processes / physiology
Cell Line
Epithelial Cells / cytology,  metabolism
Flow Cytometry
Fluorescent Dyes / chemistry
Glycoproteins / biosynthesis
Humans
Immunohistochemistry
Immunophenotyping
Male
Microscopy, Confocal
Peptides
Prostatic Hyperplasia / metabolism,  pathology*
Prostatic Neoplasms / metabolism,  pathology*
Grant Support
ID/Acronym/Agency:
G0500966//Medical Research Council
Chemical
Reg. No./Substance:
0/AC133 antigen; 0/Antigens, CD; 0/Benzimidazoles; 0/Fluorescent Dyes; 0/Glycoproteins; 0/Peptides; 23491-52-3/HOE 33342; EC 3.1.3.48/Antigens, CD45

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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