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n2:pmcid: PMC0
bibo:doi: 10.1038%2Flabinvest.2009.127
n4:contains: _:vb6614028 _:vb6613996 _:vb6614003

Subject Item: _:vb6613996
rdf:type: n4:Section
dc:title: results
n4:contains: _:vb6614000 _:vb6614001 _:vb6614002 _:vb6613997 _:vb6613998 _:vb6613999

Subject Item: _:vb6613997
rdf:type: n2:Context
rdf:value: Because anchorage-independent growth is an approximation of tumorigenesis, and CSCs are thought to be the tumor-initiating cells (>>2<<), we further tested the ability of the ALDH1A1+ and ALDH1A1− PCa cells to produce colonies by using soft agar assay.
n2:mentions: n3:11689955

Subject Item: _:vb6613998
rdf:type: n2:Context
rdf:value: The gold standard in determining CSCs is whether the testing cells can preferentially initiate tumor development in animal models (>>9<<, 22). We therefore implanted ALDH1A1+ PC3, ALDH1A1− PC3, ALDH1A1+ LNCaP, and ALDH1A1− LNCaP cells into ten NOD/SCID mice, respectively.
n2:mentions: n3:17638891

Subject Item: _:vb6613999
rdf:type: n2:Context
rdf:value: The gold standard in determining CSCs is whether the testing cells can preferentially initiate tumor development in animal models (9, >>22<<). We therefore implanted ALDH1A1+ PC3, ALDH1A1− PC3, ALDH1A1+ LNCaP, and ALDH1A1− LNCaP cells into ten NOD/SCID mice, respectively.
n2:mentions: n3:16990346

Subject Item: _:vb6614000
rdf:type: n2:Context
rdf:value: Together with previous reports that the basal cell fraction includes prostate SCs (>>5<<) and CD44+ cells represent normal SCs (5, 10), our findings of ALDH1A1+ cells comprising a subset of CD44+ cells in normal basal layer component might imply that the ALDH1A1+ cells could be enriched in SCs of prostate.
n2:mentions: n3:19040209

Subject Item: _:vb6614001
rdf:type: n2:Context
rdf:value: Together with previous reports that the basal cell fraction includes prostate SCs (5) and CD44+ cells represent normal SCs (>>5<<, 10), our findings of ALDH1A1+ cells comprising a subset of CD44+ cells in normal basal layer component might imply that the ALDH1A1+ cells could be enriched in SCs of prostate.
n2:mentions: n3:19040209

Subject Item: _:vb6614002
rdf:type: n2:Context
rdf:value: Together with previous reports that the basal cell fraction includes prostate SCs (5) and CD44+ cells represent normal SCs (5, >>10<<), our findings of ALDH1A1+ cells comprising a subset of CD44+ cells in normal basal layer component might imply that the ALDH1A1+ cells could be enriched in SCs of prostate.
n2:mentions: n3:16921491

Subject Item: _:vb6614003
rdf:type: n4:Section
dc:title: discussion
n4:contains: _:vb6614024 _:vb6614025 _:vb6614026 _:vb6614027 _:vb6614016 _:vb6614017 _:vb6614018 _:vb6614019 _:vb6614020 _:vb6614021 _:vb6614022 _:vb6614023 _:vb6614008 _:vb6614009 _:vb6614010 _:vb6614011 _:vb6614012 _:vb6614013 _:vb6614014 _:vb6614015 _:vb6614004 _:vb6614005 _:vb6614006 _:vb6614007

Subject Item: _:vb6614004
rdf:type: n2:Context
rdf:value: A Hoechst 33342 efflux assay for collecting side population (SP) that preferentially expresses drug-efflux pumps has been used to enrich CSCs of leukemia and some solid tumors (>>2<<-3). However, the approach failed to detect a reliable SP in several commonly cultured human PCa cell lines (9-10). Recently, cell surface markers have been utilized to isolate CSCs from human PCa cells. For instance, CD44+/α2ß1+ PCa cells
n2:mentions: n3:11689955 n3:16990388

Subject Item: _:vb6614005
rdf:type: n2:Context
rdf:value: However, the approach failed to detect a reliable SP in several commonly cultured human PCa cell lines (>>9<<-10). Recently, cell surface markers have been utilized to isolate CSCs from human PCa cells. For instance, CD44+/α2ß1+ PCa cells demonstrated high colony-forming efficiency and capacity to differentiate into several cell types of prostate
n2:mentions: n3:16921491 n3:17638891

Subject Item: _:vb6614006
rdf:type: n2:Context
rdf:value: For instance, CD44+/α2ß1+ PCa cells demonstrated high colony-forming efficiency and capacity to differentiate into several cell types of prostate carcinoma (>>9<<-10). However, the ability of these cells to serially reinitiate transplantable tumors has not been well documented (10). Furthermore, the reliability of using the markers to identify CSCs remains controversial (5, 10). For example,
n2:mentions: n3:16921491 n3:17638891

Subject Item: _:vb6614007
rdf:type: n2:Context
rdf:value: However, the ability of these cells to serially reinitiate transplantable tumors has not been well documented (>>10<<). Furthermore, the reliability of using the markers to identify CSCs remains controversial (5, 10). For example, Collins et al demonstrated that a subpopulation of CD44+/a2b1high/CD133+ cells in clinical PCa specimens had stem cell
n2:mentions: n3:16921491

Subject Item: _:vb6614008
rdf:type: n2:Context
rdf:value: Furthermore, the reliability of using the markers to identify CSCs remains controversial (>>5<<, 10). For example, Collins et al demonstrated that a subpopulation of CD44+/a2b1high/CD133+ cells in clinical PCa specimens had stem cell characteristics in vitro (5). However, Pfeiffer et al (23) recently used CD133 to isolate cancer
n2:mentions: n3:19040209

Subject Item: _:vb6614009
rdf:type: n2:Context
rdf:value: Furthermore, the reliability of using the markers to identify CSCs remains controversial (5, >>10<<). For example, Collins et al demonstrated that a subpopulation of CD44+/a2b1high/CD133+ cells in clinical PCa specimens had stem cell characteristics in vitro (5). However, Pfeiffer et al (23) recently used CD133 to isolate cancer stem
n2:mentions: n3:16921491

Subject Item: _:vb6614010
rdf:type: n2:Context
rdf:value: For example, Collins et al demonstrated that a subpopulation of CD44+/a2b1high/CD133+ cells in clinical PCa specimens had stem cell characteristics in vitro (>>5<<). However, Pfeiffer et al (23) recently used CD133 to isolate cancer stem cells from PCa cell lines. Only one (DU145) of 6 PCa cell lines (DU145, DuCaP, LAPC-4, 22Rv1, LNCaP, and PC-3) had detectable CD133+ population (0.01%). There was
n2:mentions: n3:19040209

Subject Item: _:vb6614011
rdf:type: n2:Context
rdf:value: In addition, although α2ß1+ PCa cells showed higher proliferative and clonogenic potentials in vitro compared with the isogenic α2ß1− PCa cells, α2ß1+ PCa cells were no more tumorigenic than the corresponding α2ß1− cells in vivo (>>9<<). Moreover, several other cell populations have been identified that share the same marker phenotype, including circulating fibrocytes (22-6), thus limiting their application in enriching CSCs from clinical specimens (24).
n2:mentions: n3:17638891

Subject Item: _:vb6614012
rdf:type: n2:Context
rdf:value: Moreover, several other cell populations have been identified that share the same marker phenotype, including circulating fibrocytes (>>22<<-6), thus limiting their application in enriching CSCs from clinical specimens (24).
n2:mentions: n3:16990346

Subject Item: _:vb6614013
rdf:type: n2:Context
rdf:value: Moreover, several other cell populations have been identified that share the same marker phenotype, including circulating fibrocytes (22-6), thus limiting their application in enriching CSCs from clinical specimens (>>24<<).
n2:mentions: n3:18182063

Subject Item: _:vb6614014
rdf:type: n2:Context
rdf:value: Previous studies strongly suggested that the basal cell layer of adult human prostate contained SCs (>>5<<, 10). For example, more than 70% proliferating cells and molecules important in maintaining fundamental properties of SC localized in this basal compartment (10, 27).
n2:mentions: n3:19040209

Subject Item: _:vb6614015
rdf:type: n2:Context
rdf:value: Previous studies strongly suggested that the basal cell layer of adult human prostate contained SCs (5, >>10<<). For example, more than 70% proliferating cells and molecules important in maintaining fundamental properties of SC localized in this basal compartment (10, 27).
n2:mentions: n3:16921491

Subject Item: _:vb6614016
rdf:type: n2:Context
rdf:value: For example, more than 70% proliferating cells and molecules important in maintaining fundamental properties of SC localized in this basal compartment (>>10<<, 27). Furthermore, less than 5% dissociated basal layer cells from adult human prostate possessed extensive proliferative capacity (27-8). In addition, under suitable culture conditions, a small population of prostatic basal cells
n2:mentions: n3:16921491

Subject Item: _:vb6614017
rdf:type: n2:Context
rdf:value: For example, more than 70% proliferating cells and molecules important in maintaining fundamental properties of SC localized in this basal compartment (10, >>27<<). Furthermore, less than 5% dissociated basal layer cells from adult human prostate possessed extensive proliferative capacity (27-8). In addition, under suitable culture conditions, a small population of prostatic basal cells maintained
n2:mentions: n3:15750631

Subject Item: _:vb6614018
rdf:type: n2:Context
rdf:value: Furthermore, less than 5% dissociated basal layer cells from adult human prostate possessed extensive proliferative capacity (>>27<<-8). In addition, under suitable culture conditions, a small population of prostatic basal cells maintained some developmental plasticity, in which, the cells transdifferentiated into neuronal/glial cells (29). Finally, CD44 was expressed
n2:mentions: n3:15750631

Subject Item: _:vb6614019
rdf:type: n2:Context
rdf:value: In addition, under suitable culture conditions, a small population of prostatic basal cells maintained some developmental plasticity, in which, the cells transdifferentiated into neuronal/glial cells (>>29<<). Finally, CD44 was expressed on most basal cells and purified CD44+ prostate epithelial cells, when co-cultured with stromal cells, produced increased amounts of PSA, probably due to the differentiation of CD44+ cells into luminal cells
n2:mentions: n3:14570561

Subject Item: _:vb6614020
rdf:type: n2:Context
rdf:value: Finally, CD44 was expressed on most basal cells and purified CD44+ prostate epithelial cells, when co-cultured with stromal cells, produced increased amounts of PSA, probably due to the differentiation of CD44+ cells into luminal cells (>>10<<, 30). CD44+ cells were therefore believed to contain normal prostate SCs and CD44 was considered as a marker for SCs in prostate epithelia (5, 10).
n2:mentions: n3:16921491

Subject Item: _:vb6614021
rdf:type: n2:Context
rdf:value: CD44 was expressed on most basal cells and purified CD44+ prostate epithelial cells, when co-cultured with stromal cells, produced increased amounts of PSA, probably due to the differentiation of CD44+ cells into luminal cells (10, >>30<<). CD44+ cells were therefore believed to contain normal prostate SCs and CD44 was considered as a marker for SCs in prostate epithelia (5, 10).
n2:mentions: n3:9380699

Subject Item: _:vb6614022
rdf:type: n2:Context
rdf:value: CD44+ cells were therefore believed to contain normal prostate SCs and CD44 was considered as a marker for SCs in prostate epithelia (>>5<<, 10). In this study, we demonstrate that a small ALDH1A1+ cell population is restricted to the CD44 positive basal layer cells of normal prostates, suggesting that the ALDH1A1+ cells in normal prostate glands might be SCs. Investigation
n2:mentions: n3:19040209

Subject Item: _:vb6614023
rdf:type: n2:Context
rdf:value: CD44+ cells were therefore believed to contain normal prostate SCs and CD44 was considered as a marker for SCs in prostate epithelia (5, >>10<<). In this study, we demonstrate that a small ALDH1A1+ cell population is restricted to the CD44 positive basal layer cells of normal prostates, suggesting that the ALDH1A1+ cells in normal prostate glands might be SCs. Investigation of
n2:mentions: n3:16921491

Subject Item: _:vb6614024
rdf:type: n2:Context
rdf:value: Furthermore, our observations also confirm concept of a hierarchical organization of cancer cells existing in PCa proposed by Tang et al and others (>>9<<, 10, 31). Based on the concept, CSCs could derive from mutated CSs, and PCa cells are organized as a hierarchy with CSCs sitting at the apex and having the ability to develop (or differentiate) into a spectrum of more mature progeny. The
n2:mentions: n3:17638891

Subject Item: _:vb6614025
rdf:type: n2:Context
rdf:value: Furthermore, our observations also confirm concept of a hierarchical organization of cancer cells existing in PCa proposed by Tang et al and others (9, >>10<<, 31). Based on the concept, CSCs could derive from mutated CSs, and PCa cells are organized as a hierarchy with CSCs sitting at the apex and having the ability to develop (or differentiate) into a spectrum of more mature progeny. The
n2:mentions: n3:16921491

Subject Item: _:vb6614026
rdf:type: n2:Context
rdf:value: Furthermore, our observations also confirm concept of a hierarchical organization of cancer cells existing in PCa proposed by Tang et al and others (9, 10, >>31<<). Based on the concept, CSCs could derive from mutated CSs, and PCa cells are organized as a hierarchy with CSCs sitting at the apex and having the ability to develop (or differentiate) into a spectrum of more mature progeny. The
n2:mentions: n3:17510410

Subject Item: _:vb6614027
rdf:type: n2:Context
rdf:value: There are few markers clinically available for accurate prediction of prognosis of PCa patients besides PSA and Gleason sum (>>32<<). Because of the heterogeneous nature of PCa, it has become clear that a combination of accurate prognostic indicators is needed to improve the ability for identifying aggressive disease, so that more individualized treatments could be
n2:mentions: n3:16557596

Subject Item: _:vb6614028
rdf:type: n4:Section
dc:title: material and methods
n4:contains: _:vb6614036 _:vb6614032 _:vb6614033 _:vb6614034 _:vb6614035 _:vb6614029 _:vb6614030 _:vb6614031

Subject Item: _:vb6614029
rdf:type: n2:Context
rdf:value: An Aldefluor kit (StemCell Technologies, Durham, NC) optimized for interaction with human ALDH1A1 was used to identify ALDH1A1+ cells as previously described (>>18<<). Briefly, the brightly fluorescent ALDH1A1 expressing cells were detected by using an Aria cell sorter (BD Biosciences, San Jose, CA).
n2:mentions: n3:19276181

Subject Item: _:vb6614030
rdf:type: n2:Context
rdf:value: Cells were plated at density of 200 per well in a six-well tissue culture dish, as described in our previous report (>>18<<). Clones with >50 cells were scored at the end of week two. The percentage of cells that initiated a clone was presented as cloning efficiency.
n2:mentions: n3:19276181

Subject Item: _:vb6614031
rdf:type: n2:Context
rdf:value: For clonogenic assays, cells were plated at 1,000 per well in six-well culture dishes coated with a thin layer of 1% solidified agar (>>9<<, 18). Spheres that arose within three weeks were considered as clonogenicity. For each cell type, triplicate samples were performed and the clones or spheres were counted by two individuals in a blind fashion.
n2:mentions: n3:17638891

Subject Item: _:vb6614032
rdf:type: n2:Context
rdf:value: For clonogenic assays, cells were plated at 1,000 per well in six-well culture dishes coated with a thin layer of 1% solidified agar (9, >>18<<). Spheres that arose within three weeks were considered as clonogenicity. For each cell type, triplicate samples were performed and the clones or spheres were counted by two individuals in a blind fashion.
n2:mentions: n3:19276181

Subject Item: _:vb6614033
rdf:type: n2:Context
rdf:value: The cells were then implanted into the dorsal prostates of ten NOD/SCID mice, which has been widely used as an ‘‘orthotopic’’ implantation site for human PCa (>>9<<). Tumorigenicity was measured mainly by tumor incidence (i.e., the number of tumors/number of injections), size of tumor, and latency (i.e., time from injection to detection of palpable tumors). The mice were observed for six weeks to
n2:mentions: n3:17638891

Subject Item: _:vb6614034
rdf:type: n2:Context
rdf:value: A part of each engrafted tumor tissue was dissociated with collagenase IV (Sigma) and incubated with mechanical disruption as previously described (>>18<<). Serial transplantations of the PCa xenografts were performed by regrafting the freshly disassociated cells into male mice. The disassociated cells were also reanalyzed by using Aldefluor assay for FACS analysis as described above.
n2:mentions: n3:19276181

Subject Item: _:vb6614035
rdf:type: n2:Context
rdf:value: by using antibodies against ALDH1A1 (Santa Cruz Biotechnology, Santa Cruz, CA), CD44 (eBioscience, San Diego, CA), AR (Santa Cruz Biotechnology), and chromogranin A (Vector Laboratories, Inc., Burlingame, CA) as previously described (>>20<<-21). Immunoreactive staining intensity for each antibody was rated according to the following scale:
n2:mentions: n3:19189306 n3:19058139

Subject Item: _:vb6614036
rdf:type: n2:Context
rdf:value: Buffalo, NY), as previously described (>>18<<). Each batch of slides contained a positive control and a negative control.
n2:mentions: n3:19276181

Subject Item: _:vb420926807
rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 49
n2:hasRelevantPaperId: n3:18371393

Subject Item: _:vb420926808
rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 30
n2:hasRelevantPaperId: n3:19276181

Subject Item: _:vb420926809
rdf:type: n2:RelevantBibliographicResource
n2:RelevantScore: 25
n2:hasRelevantPaperId: n3:12629218

Subject Item: _:vb420926810
rdf:type: n2:RelevantBibliographicResource
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