Mutation within the adult human stem cell (SC) compartment has been proposed as a factor in the initiation and promotion of carcinogenesis. Isolation of these cancer stem cells (CSCs) has proven difficult, limiting their subsequent phenotypic, functional, and genetic characterization. We have used the Hoechst 33342 dye efflux technique to isolate an epithelial side population (SP) from genitourinary (GU) cancers, which is enriched for cells with SC traits. With informed consent, samples were taken from patients with primary tumors and undergoing surgery for prostatic (CaP), invasive bladder transitional cell (TCC), and renal cell carcinomas (RCC). Single cell epithelial suspensions were extracted from these and incubated with Hoechst 33342. Hoechst SP/non-SP profiles were then generated by flow cytometry using standardized protocols. SP/non-SP cell cycle status was established by Hoechst 33342 and Pyronin Y staining. Immunocytochemistry staining was performed for markers suggested as stem markers as well as lineage-specific markers. Functionality was determined using colony-forming assays and long-term monolayer culture. A characteristic verapamil-sensitive SP was isolated from all 3 urological malignancies and represented 0.57% +/- 0.11% (CaP), 0.52% +/- 0.49% (TCC), and 5.9% +/- 0.9% (RCC) of the total epithelial population. Cell cycle analysis showed that the SP had enhanced numbers of cells in G(0) as compared to the total cell population (CaP 12.4% +/- 3.2 vs. 3.8% +/- 1.0, RCC 23.2% +/- 3.4 vs. 1.8% +/- 0.9, and TCC 28.5% +/- 4.9 vs. 4% +/- 1.3). Immunocytochemistry demonstrated an increased expression of proliferative and putative stem markers within the SP fraction. Cultures confirmed significant enhancement of colony-forming ability and proliferative capacity of the SP fraction. A characteristic SP enriched for stem-like cells has been isolated from the 3 most common urological malignancies. This provides strong evidence that Hoechst 33342 efflux is a conserved and unified mechanism in GU cancer.

You have accessJournal of Urology1 Apr 2009GENETIC PROFILING OF THE STEM CELL ENRICHED PROSTATE SIDE POPULATION Benjamin R Grey, Jeremy E Oates, Claire A Hart, Yvonne Hey, Gill Newton, Sian Dibben, Stuart D Pepper, Michael D Brown, and Noel W Clarke Benjamin R GreyBenjamin R Grey More articles by this author , Jeremy E OatesJeremy E Oates More articles by this author , Claire A HartClaire A Hart More articles by this author , Yvonne HeyYvonne Hey More articles by this author , Gill NewtonGill Newton More articles by this author , Sian DibbenSian Dibben More articles by this author , Stuart D PepperStuart D Pepper More articles by this author , Michael D BrownMichael D Brown More articles by this author , and Noel W ClarkeNoel W Clarke More articles by this author View All Author Informationhttps://doi.org/10.1016/S0022-5347(09)60132-4AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail "GENETIC PROFILING OF THE STEM CELL ENRICHED PROSTATE SIDE POPULATION." The Journal of Urology, 181(4S), pp. 42–43 © 2009 by American Urological AssociationFiguresReferencesRelatedDetails Volume 181 Issue 4S April 2009 Page: 42-43 Advertisement Copyright & Permissions© 2009 by American Urological AssociationMetrics Author Information Benjamin R Grey More articles by this author Jeremy E Oates More articles by this author Claire A Hart More articles by this author Yvonne Hey More articles by this author Gill Newton More articles by this author Sian Dibben More articles by this author Stuart D Pepper More articles by this author Michael D Brown More articles by this author Noel W Clarke More articles by this author Expand All Advertisement PDF downloadLoading ...

What do we conclude from this? While being a term that has developed, radical nephrectomy is probably a reasonable phrase for removal of the kidney secondary to malignancy (with or without adrenalectomy and lymphadenectomy). However, simple nephrectomy is a misnomer, as ‘simple’ nephrectomies are not, on average, easier than radical nephrectomies. Furthermore, complication rates are comparable if not greater than nephrectomies for malignancy. This terminology could be particularly troublesome should a case proceed to the medicolegal arena, in which, e.g. one could be criticised for complications in a ‘simple’ nephrectomy case? We thus argue that the term ‘simple’ nephrectomy be abandoned and replaced by the more appropriate terminology of ‘non-radical’ nephrectomy, to emphasize surgical removal of the kidney for nonmalignant conditions.