26S Proteasome Activity Is Down-Regulated in Lung Cancer Stem-Like Cells Propagated In Vitro
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{"title"=>"26S proteasome activity is down-regulated in lung cancer stem-like cells propagated In Vitro", "type"=>"journal", "authors"=>[{"first_name"=>"Jing", "last_name"=>"Pan", "scopus_author_id"=>"56335049500"}, {"first_name"=>"Qi", "last_name"=>"Zhang", "scopus_author_id"=>"55142767800"}, {"first_name"=>"Yian", "last_name"=>"Wang", "scopus_author_id"=>"7601502017"}, {"first_name"=>"Ming", "last_name"=>"You", "scopus_author_id"=>"7102487803"}], "year"=>2010, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"20949018", "doi"=>"10.1371/journal.pone.0013298", "sgr"=>"78149423083", "isbn"=>"1932-6203", "scopus"=>"2-s2.0-78149423083", "issn"=>"19326203", "pui"=>"359930893"}, "id"=>"b3251776-52e5-3b6e-b7bd-ea077ff11c70", "abstract"=>"Cancer stem cells (CSCs) are a small subset of cancer cells capable of self-renewal and tumor maintenance. Eradicating cancer stem cells, the root of tumor origin and recurrence, has emerged as one promising approach to improve lung cancer survival. Cancer stem cells are reported to reside in the side population (SP) of cultured lung cancer cells. We report here the coexistence of a distinct population of non-SP (NSP) cells that have equivalent self-renewal capacity compared to SP cells in a lung tumor sphere assay. Compared with the corresponding cells in monolayer cultures, lung tumor spheres, formed from human non-small cell lung carcinoma cell lines A549 or H1299, showed marked morphologic differences and increased expression of the stem cell markers CD133 and OCT3/4. Lung tumor spheres also exhibited increased tumorigenic potential as only 10,000 lung tumor sphere cells were required to produce xenografts tumors in nude mice, whereas the same number of monolayer cells failed to induce tumors. We also demonstrate that lung tumor spheres showed decreased 26S proteasome activity compared to monolayer. By using the ZsGreen-cODC (C-terminal sequence that directs degradation of Ornithine Decarboxylase) reporter assay in NSCLC cell lines, only less than 1% monolayer cultures were ZsGreen positive indicating low 26S proteasome, whereas lung tumor sphere showed increased numbers of ZsGreen-positive cells, suggesting the enrichment of CSCs in sphere cultures.", "link"=>"http://www.mendeley.com/research/26s-proteasome-activity-downregulated-lung-cancer-stemlike-cells-propagated-vitro", "reader_count"=>36, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>4, "Student > Doctoral Student"=>3, "Researcher"=>7, "Student > Ph. D. Student"=>10, "Student > Master"=>8, "Student > Bachelor"=>1, "Professor"=>2, "Student > Postgraduate"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>4, "Student > Doctoral Student"=>3, "Researcher"=>7, "Student > Ph. D. Student"=>10, "Student > Master"=>8, "Student > Bachelor"=>1, "Professor"=>2, "Student > Postgraduate"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>5, "Agricultural and Biological Sciences"=>20, "Medicine and Dentistry"=>8, "Veterinary Science and Veterinary Medicine"=>1, "Chemistry"=>1, "Social Sciences"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>8}, "Chemistry"=>{"Chemistry"=>1}, "Social Sciences"=>{"Social Sciences"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>20}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Veterinary Science and Veterinary Medicine"=>{"Veterinary Science and Veterinary Medicine"=>1}}, "reader_count_by_country"=>{"United States"=>1, "South Africa"=>1, "Spain"=>1}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/825916"], "description"=>"<p>Tumorigenic potential of tumor sphere cells is greater than monolayer cancer cells. Monolayer and tumor sphere cells from A549 were injected s.c. into athymic nude mice at concentrations of 1×10<sup>4</sup> cells/ml. Mice were sacrificed at day 60, and tumor was surgically removed and measured, and the mean values of the tumor size were shown in the table.</p>", "links"=>[], "tags"=>["spheres", "tumorigenic"], "article_id"=>496286, "categories"=>["Cell Biology", "Cancer"], "users"=>["Jing Pan", "Qi Zhang", "Yian Wang", "Ming You"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0013298.t001", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Lung_tumor_spheres_have_high_tumorigenic_potential_/496286", "title"=>"Lung tumor spheres have high tumorigenic potential.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 23:01:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/825377"], "description"=>"<p><b>A.</b> Lung tumor spheres showed increased CD133 expression. Expression of CD44 and CD133 in monolayer and tumor sphere cells, as determined by two-color flow cytometry analysis. Isotype-matched monoclonal antibodies and a preimmune second antiserum were used as negative controls. The percentage of single- and double-positive cells are indicated. <b>B.</b> Lung tumor spheres showed increased OCT3/4 expression. An average fluorescence intensity of OCT3/4 in control (grey area), monolayer cells (dotted line) and tumor spheres (solid line) is shown. Percentages of positive cells are indicated in the table.</p>", "links"=>[], "tags"=>["spheres", "cancer"], "article_id"=>495755, "categories"=>["Cell Biology", "Cancer"], "users"=>["Jing Pan", "Qi Zhang", "Yian Wang", "Ming You"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0013298.g002", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Lung_tumor_spheres_exhibit_cancer_stem_cell_features_/495755", "title"=>"Lung tumor spheres exhibit cancer stem cell features.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 22:58:31"}
  • {"files"=>["https://ndownloader.figshare.com/files/825521"], "description"=>"<p>Surgically removed tumor was fixed in buffered formalin and subsequently analyzed by immunohistochemistry (IHC) by standard protocol.</p>", "links"=>[], "tags"=>["engrafted", "tumors"], "article_id"=>495887, "categories"=>["Cell Biology", "Cancer"], "users"=>["Jing Pan", "Qi Zhang", "Yian Wang", "Ming You"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0013298.g003", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Histopathologic_examination_of_the_engrafted_tumors_by_H_amp_E_and_946_Catenin_staining_/495887", "title"=>"Histopathologic examination of the engrafted tumors by H&E and β-Catenin staining.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 22:59:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/825710"], "description"=>"<p><b>A.</b> caspase-like, <b>B.</b> chymotrypsin-like, and <b>C.</b> trypsin-like activity of monolayer culture and tumor spheres from A549 and H1299 cells was measued over the indicated times. <b>D.</b> End point analysis (120 min) of 26S proteasome activities in monolayer cultures and tumor spheres. Mean ± SD is plotted and derived from three independent experiments. Student's paired, and two-tailed t-tests were performed, *p<0.05, **p<0.01, and ***p<0.001 (three replicates per experiment).</p>", "links"=>[], "tags"=>["proteasome", "activities", "26s", "a549", "h1299"], "article_id"=>496080, "categories"=>["Cell Biology", "Cancer"], "users"=>["Jing Pan", "Qi Zhang", "Yian Wang", "Ming You"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0013298.g005", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Decreased_proteasome_activities_of_the_26S_proteasome_in_A549_and_H1299_tumor_spheres_/496080", "title"=>"Decreased proteasome activities of the 26S proteasome in A549 and H1299 tumor spheres.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 23:00:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/825643"], "description"=>"<p><b>A.</b> cisplatin, <b>B.</b> doxorubicin resistance of monolayer culture and tumor spheres were analyzed. Cells dissociated from lung tumor spheres or monolayers were plated out in 96-well plate at 2×10<sup>4</sup> cells/ml, 24 h later, media containing different concentrations of cisplatin or doxorubicin was added. After 48 h culturing, 10% Alamar Blue was added and read at 544/590 nm, the percent viability was calculated relative to cells not exposed to any chemotherapeutic drugs. The results represent the means ± SD.</p>", "links"=>[], "tags"=>["monolayer"], "article_id"=>496011, "categories"=>["Cell Biology", "Cancer"], "users"=>["Jing Pan", "Qi Zhang", "Yian Wang", "Ming You"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0013298.g004", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Chemoresistence_of_monolayer_and_tumor_sphere_/496011", "title"=>"Chemoresistence of monolayer and tumor sphere.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 22:59:53"}
  • {"files"=>["https://ndownloader.figshare.com/files/825256"], "description"=>"<p><b>A.</b> Characteristic Hoechst 33342 dye staining profile demonstrating the existence of SP (polygon gated) and NSP (ellipse gated) cells in human A549 and H1299 adenocarcinoma cells. Percentage of SP cells is indicated. Cells were stained with 5 mg/ml Hoechst33342 (HO), and control cells were also co-stained with 10 mM fumitremorgin C (FTC) to specify the SP cells. Cells were sorted using a Beckman MoFlo cytometer and data were annotated using FCS Express. <b>B.</b> Phase-contrast photographs of lung tumor spheres obtained from SP and NSP cells. SP and NSP cells (1000 cell/ml) were plated onto ultra low adherent flasks in serum free media supplemented with growth factors and cultivated as described in Material and <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013298#s2\" target=\"_blank\">Methods</a>. <b>C.</b> Re-stain of SP and NSP cells from A549 and H1299 with Hoechst 33342 dye. Percentage of SP cells is labeled.</p>", "links"=>[], "tags"=>["nsp", "cells", "spheres", "adenocarcinoma"], "article_id"=>495626, "categories"=>["Cell Biology", "Cancer"], "users"=>["Jing Pan", "Qi Zhang", "Yian Wang", "Ming You"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0013298.g001", "stats"=>{"downloads"=>1, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_SP_and_NSP_cells_form_tumor_spheres_with_equal_capacity_in_human_adenocarcinoma_cell_lines_/495626", "title"=>"SP and NSP cells form tumor spheres with equal capacity in human adenocarcinoma cell lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 22:57:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/825818"], "description"=>"<p><b>A.</b> Frequency of ZsGreen cells in A549 and H1299 monolayer cultures. <b>B.</b> Frequency of ZsGreen cells in A549 and H12199-derived tumor spheres with accumulation of ZsGreen-cODC, and thus low proteasome activity is indicated. <b>C.</b> Number of spheres formed from the ZsGreen positive and the ZsGreen negative cells after sorting with flow cytometry into 96-well plates. ZsGree-positive and –negative cells were sorted into 96-well plate (100cells/well), after cultured in tumor sphere media for 2 weeks; number of formed spheres was counted. Student's paired, and two-tailed t-tests were performed, ***p<0.001 (three replicates per experiment).</p>", "links"=>[], "tags"=>["cscs", "proteasome"], "article_id"=>496193, "categories"=>["Cell Biology", "Cancer"], "users"=>["Jing Pan", "Qi Zhang", "Yian Wang", "Ming You"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0013298.g006", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Track_CSCs_in_tumor_sphere_based_on_their_proteasome_activity_/496193", "title"=>"Track CSCs in tumor sphere based on their proteasome activity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 23:00:49"}

PMC Usage Stats | Further Information

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Relative Metric

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