Human α2β1HI CD133+VE Epithelial Prostate Stem Cells Express Low Levels of Active Androgen Receptor
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{"title"=>"Human α2β1HI CD133+VE Epithelial Prostate Stem Cells Express Low Levels of Active Androgen Receptor", "type"=>"journal", "authors"=>[{"first_name"=>"Stuart C.", "last_name"=>"Williamson", "scopus_author_id"=>"54394602700"}, {"first_name"=>"Anastasia C.", "last_name"=>"Hepburn", "scopus_author_id"=>"55470017100"}, {"first_name"=>"Laura", "last_name"=>"Wilson", "scopus_author_id"=>"56420413900"}, {"first_name"=>"Kelly", "last_name"=>"Coffey", "scopus_author_id"=>"55385739600"}, {"first_name"=>"Claudia A.", "last_name"=>"Ryan-Munden", "scopus_author_id"=>"55386004600"}, {"first_name"=>"Deepali", "last_name"=>"Pal", "scopus_author_id"=>"55471444900"}, {"first_name"=>"Hing Y.", "last_name"=>"Leung", "scopus_author_id"=>"7202811500"}, {"first_name"=>"Craig N.", "last_name"=>"Robson", "scopus_author_id"=>"7005113083"}, {"first_name"=>"Rakesh", "last_name"=>"Heer", "scopus_author_id"=>"8272209200"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-84868699649", "pui"=>"366025076", "doi"=>"10.1371/journal.pone.0048944", "issn"=>"19326203", "pmid"=>"23145034", "sgr"=>"84868699649"}, "id"=>"b70c16fd-302d-3352-889c-08d0dc399c3d", "abstract"=>"Stem cells are thought to be the cell of origin in malignant transformation in many tissues, but their role in human prostate carcinogenesis continues to be debated. One of the conflicts with this model is that cancer stem cells have been described to lack androgen receptor (AR) expression, which is of established importance in prostate cancer initiation and progression. We re-examined the expression patterns of AR within adult prostate epithelial differentiation using an optimised sensitive and specific approach examining transcript, protein and AR regulated gene expression. Highly enriched populations were isolated consisting of stem (α(2)β(1)(HI) CD133(+VE)), transiently amplifying (α(2)β(1)(HI) CD133(-VE)) and terminally differentiated (α(2)β(1)(LOW) CD133(-VE)) cells. AR transcript and protein expression was confirmed in α(2)β(1)(HI) CD133(+VE) and CD133(-VE) progenitor cells. Flow cytometry confirmed that median (±SD) fraction of cells expressing AR were 77% (±6%) in α(2)β(1)(HI) CD133(+VE) stem cells and 68% (±12%) in α(2)β(1)(HI) CD133(-VE) transiently amplifying cells. However, 3-fold lower levels of total AR protein expression (peak and median immunofluorescence) were present in α(2)β(1)(HI) CD133(+VE) stem cells compared with differentiated cells. This finding was confirmed with dual immunostaining of prostate sections for AR and CD133, which again demonstrated low levels of AR within basal CD133(+VE) cells. Activity of the AR was confirmed in prostate progenitor cells by the expression of low levels of the AR regulated genes PSA, KLK2 and TMPRSS2. The confirmation of AR expression in prostate progenitor cells allows integration of the cancer stem cell theory with the established models of prostate cancer initiation based on a functional AR. Further study of specific AR functions in prostate stem and differentiated cells may highlight novel mechanisms of prostate homeostasis and insights into tumourigenesis.", "link"=>"http://www.mendeley.com/research/human-%CE%B12%CE%B21hi-cd133ve-epithelial-prostate-stem-cells-express-low-levels-active-androgen-receptor", "reader_count"=>3, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Medicine and Dentistry"=>1, "Agricultural and Biological Sciences"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>1}, "Unspecified"=>{"Unspecified"=>1}}, "reader_count_by_country"=>{"United Kingdom"=>1}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/546734"], "description"=>"<p><b>A)</b> Percentage of cells staining above a no antibody control for either isotype antibody (hollow points) or PG-21 AR antibody (solid points) in LNCaP (circles) or PC3 (triangles) across a dilution series. <b>B)</b> Representative staining patterns for PC3 (upper dotplots) and LNCaP (lower dotplots) for either 1∶200 isotype antibody (left dotplots) or 1∶200 PG-21 AR antibody (right dotplots) of equivalent concentrations. Gates set according to isotype control. <b>C)</b> Left dotplot representative of staining of LNCaP with isotype control, right dotplots representative of AR staining in non-transfected LNCaP (upper), LNCaP transfected with scrambled siRNA (middle dotplot) and LNCaP transfected with AR siRNA (lower). Gates were set according to an appropriate isotype control. <b>D)</b> Percentage of cells staining positive for AR relative to an isotype control in non-transfected LNCaP, LNCaP transfected with scrambled siRNA and LNCaP transfected with AR siRNA. Error bars represent standard error of the mean for n = 3. <b>E)</b> Western blots of AR expression for non-transfected LNCaP, LNCaP transfected with scrambled siRNA and LNCaP transfected with AR siRNA are shown using a different AR antibody (G122-434, BD Pharmingen).</p>", "links"=>[], "tags"=>["ar", "detection"], "article_id"=>217224, "categories"=>["Cancer", "Cell Biology", "Developmental Biology"], "users"=>["Stuart C. Williamson", "Anastasia C. Hepburn", "Laura Wilson", "Kelly Coffey", "Claudia A. Ryan-Munden", "Deepali Pal", "Hing Y. Leung", "Craig N. Robson", "Rakesh Heer"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0048944.g002", "stats"=>{"downloads"=>3, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Validation_of_AR_detection_with_flow_cytometry_/217224", "title"=>"Validation of AR detection with flow cytometry.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 20:39:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/546553"], "description"=>"<p><b>A)</b> Schematic work flow for enrichment of epithelial cells for assessment of AR expression. <b>B)</b> Purity of selection by expression of the lineage specific markers CD24 (epithelial), CD146 (endothelial) and CD45 (haematopoietic) normalised to GAPDH following real-time PCR for unsorted prostate epithelia and EpCAM/HEA sorted epithelia, error bars represent standard error of the mean for n = 3. <b>C)</b> CD133/1 Sorted samples were assessed for purity by co-expression of the CD133/2 epitope, confirming that these two epitopes are co-expressed in the prostate and that our CD133 selection efficiently enriches for CD133<sup>+VE</sup> cells: Upper dotplot representative of CD133/2 staining for unsorted α<sub>2</sub>β<sub>1</sub><sup>HI</sup> epithelial cells; lower left dotplot representative of CD133/2 staining for α<sub>2</sub>β<sub>1</sub><sup>HI</sup> CD133/1<sup>–VE</sup> cells; lower right dotplot representative of CD133/2 staining for α<sub>2</sub>β<sub>1</sub><sup>HI</sup> CD133/1<sup>+VE</sup> cells. Gates are set according to appropriate isotype controls. <b>D)</b> Confirmation of CD133 enrichment with real-time PCR. CD133 expression is shown normalised to GAPDH, error bars represent standard error of the mean n = 10.</p>", "links"=>[], "tags"=>["enrichment"], "article_id"=>217042, "categories"=>["Cancer", "Cell Biology", "Developmental Biology"], "users"=>["Stuart C. Williamson", "Anastasia C. Hepburn", "Laura Wilson", "Kelly Coffey", "Claudia A. Ryan-Munden", "Deepali Pal", "Hing Y. Leung", "Craig N. Robson", "Rakesh Heer"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0048944.g001", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Strategy_of_enrichment_for_required_cell_types_/217042", "title"=>"Strategy of enrichment for required cell types.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 20:38:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/547199"], "description"=>"<p>Red stain represents AR expression, blue stain represents CD133 expression, black bars are 50 µm scale bars. A) Representative image of AR staining within prostatic acini. Red arrow identifies basal layer, blue arrow identifies luminal layer. B) Representative image of CD133 and AR expression in the prostate epithelium, insert shows CD133<sup>+VE</sup> AR <sup>+VE</sup> cell within the basal layer.</p>", "links"=>[], "tags"=>["ar", "dual"], "article_id"=>217687, "categories"=>["Cancer", "Cell Biology", "Developmental Biology"], "users"=>["Stuart C. Williamson", "Anastasia C. Hepburn", "Laura Wilson", "Kelly Coffey", "Claudia A. Ryan-Munden", "Deepali Pal", "Hing Y. Leung", "Craig N. Robson", "Rakesh Heer"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0048944.g005", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_CD133_and_AR_dual_immunohistochemistry_/217687", "title"=>"CD133 and AR dual immunohistochemistry.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 20:41:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/547031"], "description"=>"<p>A) Representative images of expression of CD133 and AR counterstained with DRAQ5™ within prostate EpCAM<sup>+VE</sup> α<sub>2</sub>β<sub>1</sub><sup>HI</sup> CD133<sup>+VE</sup> (Left panel), α<sub>2</sub>β<sub>1</sub><sup>HI</sup> CD133<sup>–VE</sup> (central panel) and α<sub>2</sub>β<sub>1</sub><sup>LOW</sup> CD133<sup>–VE</sup> cells (right panel). <b>B)</b> Expression of the AR regulated genes PSA, KLK2 and TMPRSS2 normalised to GAPDH (n = 10) (p<0.001 comparing α<sub>2</sub>β<sub>1</sub><sup>HI</sup> and α<sub>2</sub>β<sub>1</sub><sup>LOW</sup>). Error bars represent standard error of the mean.</p>", "links"=>[], "tags"=>["prostate", "epithelial", "hierarchy"], "article_id"=>217518, "categories"=>["Cancer", "Cell Biology", "Developmental Biology"], "users"=>["Stuart C. Williamson", "Anastasia C. Hepburn", "Laura Wilson", "Kelly Coffey", "Claudia A. Ryan-Munden", "Deepali Pal", "Hing Y. Leung", "Craig N. Robson", "Rakesh Heer"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0048944.g004", "stats"=>{"downloads"=>3, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_AR_expression_and_activity_within_the_prostate_epithelial_hierarchy_of_differentiation_/217518", "title"=>"AR expression and activity within the prostate epithelial hierarchy of differentiation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 20:40:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/546876"], "description"=>"<p>Error bars represent standard error of the mean. <b>A)</b> Expression of AR transcript relative to GAPDH. Error bars represent standard error of the mean for n = 10. <b>B)</b> Upper dotplot representative of CD133 staining for progenitor α<sub>2</sub>β<sub>1</sub><sup>HI</sup> cells. Lower left dotplot representative of AR expression in CD133<sup>–VE</sup> gated α<sub>2</sub>β<sub>1</sub><sup>HI</sup> cells. Lower right dotplot representative of AR expression in CD133<sup>+VE</sup> gated α<sub>2</sub>β<sub>1</sub><sup>HI</sup> cells. Gates were set according to appropriate isotype controls. <b>C)</b> Mean percentage of cells expressing the AR in CD133<sup>+VE</sup> and CD133<sup>–VE</sup> α<sub>2</sub>β<sub>1</sub><sup>HI</sup> cells (n = 6). <b>D)</b> Representative histograms for fluorescence of α<sub>2</sub>β<sub>1</sub><sup>HI</sup> and α<sub>2</sub>β<sub>1</sub><sup>LOW</sup> isotype controls and AR detection. <b>E)</b> Mean fold change in median staining relative to isotype control for AR stained α<sub>2</sub>β<sub>1</sub><sup>HI</sup> cells and α<sub>2</sub>β<sub>1</sub><sup>LOW</sup> cells (n = 6).</p>", "links"=>[], "tags"=>["ar", "prostate", "epithelial", "hierarchy"], "article_id"=>217359, "categories"=>["Cancer", "Cell Biology", "Developmental Biology"], "users"=>["Stuart C. Williamson", "Anastasia C. Hepburn", "Laura Wilson", "Kelly Coffey", "Claudia A. Ryan-Munden", "Deepali Pal", "Hing Y. Leung", "Craig N. Robson", "Rakesh Heer"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0048944.g003", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_of_the_AR_within_the_prostate_epithelial_hierarchy_of_differentiation_/217359", "title"=>"Expression of the AR within the prostate epithelial hierarchy of differentiation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 20:40:06"}

PMC Usage Stats | Further Information

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Relative Metric

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