Epithelial Mesenchymal Transition and Pancreatic Tumor Initiating CD44+/EpCAM+ Cells Are Inhibited by γ-Secretase Inhibitor IX
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{"title"=>"Epithelial Mesenchymal Transition and Pancreatic Tumor Initiating CD44+/EpCAM+ Cells Are Inhibited by γ-Secretase Inhibitor IX", "type"=>"journal", "authors"=>[{"first_name"=>"Vindhya", "last_name"=>"Palagani", "scopus_author_id"=>"36462675800"}, {"first_name"=>"Mona", "last_name"=>"El Khatib", "scopus_author_id"=>"55229454100"}, {"first_name"=>"Uta", "last_name"=>"Kossatz", "scopus_author_id"=>"6506463563"}, {"first_name"=>"Przemyslaw", "last_name"=>"Bozko", "scopus_author_id"=>"6507858927"}, {"first_name"=>"Martin R.", "last_name"=>"Müller", "scopus_author_id"=>"7404688936"}, {"first_name"=>"Michael P.", "last_name"=>"Manns", "scopus_author_id"=>"36479131800"}, {"first_name"=>"Till", "last_name"=>"Krech", "scopus_author_id"=>"25224739200"}, {"first_name"=>"Nisar P.", "last_name"=>"Malek", "scopus_author_id"=>"6701536363"}, {"first_name"=>"Ruben R.", "last_name"=>"Plentz", "scopus_author_id"=>"6602207506"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"doi"=>"10.1371/journal.pone.0046514", "issn"=>"19326203", "sgr"=>"84867687369", "scopus"=>"2-s2.0-84867687369", "pui"=>"365883748", "pmid"=>"23094026", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)"}, "id"=>"3b3a0081-8c36-3dd8-84cb-4400158d84d3", "abstract"=>"Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease with a high rate of metastasis. Recent studies have indicated that the Notch signalling pathway is important in PDAC initiation and maintenance, although the specific cell biological roles of the pathway remain to be established. Here we sought to examine this question in established pancreatic cancer cell lines using the γ-secretase inhibitor IX (GSI IX) to inactivate Notch. Based on the known roles of Notch in development and stem cell biology, we focused on effects on epithelial mesenchymal transition (EMT) and on pancreatic tumor initiating CD44+/EpCAM+ cells. We analyzed the effect of the GSI IX on growth and epithelial plasticity of human pancreatic cancer cell lines, and on the tumorigenicity of pancreatic tumor initiating CD44+/EpCAM+ cells. Notably, apoptosis was induced after GSI IX treatment and EMT markers were selectively targeted. Furthermore, under GSI IX treatment, decline in the growth of pancreatic tumor initiating CD44+/EpCAM+ cells was observed in vitro and in a xenograft mouse model. This study demonstrates a central role of Notch signalling pathway in pancreatic cancer pathogenesis and identifies an effective approach to inhibit selectively EMT and suppress tumorigenesis by eliminating pancreatic tumor initiating CD44+/EpCAM+ cells.", "link"=>"http://www.mendeley.com/research/epithelial-mesenchymal-transition-pancreatic-tumor-initiating-cd44epcam-cells-inhibited-%CE%B3secretase-i", "reader_count"=>22, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>9, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>4, "Student > Postgraduate"=>1, "Student > Master"=>2, "Student > Bachelor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>9, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>4, "Student > Postgraduate"=>1, "Student > Master"=>2, "Student > Bachelor"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>6, "Agricultural and Biological Sciences"=>7, "Medicine and Dentistry"=>7, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Computer Science"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>7}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>7}, "Computer Science"=>{"Computer Science"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>6}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"Canada"=>1, "Finland"=>1, "France"=>1}, "group_count"=>2}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/555004"], "description"=>"<p>Treatment with GSI IX suppresses the migration potential of the human pancreatic cancer cell lines KP3 and BxPC3. Wound healing experiments of (A) KP3 and (B) BxPC3 cells cultured with GSI (2.5 µM, 5 µM, 10 µM) or control (DMSO). A scratch was made at (time 0 h) in both KP3 and BxPC3 and maintained for 24 h in conditioned medium with GSI or DMSO. The dotted lines are representing the edges of the wound. Photographs were taken under light microscope (10× magnification). After 24 h (A) Kp3 and (B) BxPC3 showed significant inhibition under 5 and 10 µM GSI treatment. In DMSO treated cells 80% to 90% of the wound healing was observed after 24 hrs. (C,D) The migration index was calculated as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046514#s2\" target=\"_blank\">Material and Methods</a> and plotted in bar graphs. P values were calculated with ANOVA analysis of variance along with Bonferroni post test. The error bar represents standard deviation. Differences were considered as statistically significant when the P-value was less <0.05 and non significant “n.s.” when the P-value was higher >0.05. The error bar represents standard deviation.</p>", "links"=>[], "tags"=>["pivotal", "pancreatic", "cancer"], "article_id"=>225488, "categories"=>["Cancer", "Chemistry", "Cell Biology"], "users"=>["Vindhya Palagani", "Mona El Khatib", "Uta Kossatz", "Przemyslaw Bozko", "Martin R. Müller", "Michael P. Manns", "Till Krech", "Nisar P. Malek", "Ruben R. Plentz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0046514.g002", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Notch_plays_a_pivotal_role_for_the_regulation_of_migration_in_human_pancreatic_cancer_cells_/225488", "title"=>"Notch plays a pivotal role for the regulation of migration in human pancreatic cancer cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-10-19 01:31:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/295394", "https://ndownloader.figshare.com/files/295430", "https://ndownloader.figshare.com/files/295497", "https://ndownloader.figshare.com/files/295537"], "description"=>"<div><p>Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease with a high rate of metastasis. Recent studies have indicated that the Notch signalling pathway is important in PDAC initiation and maintenance, although the specific cell biological roles of the pathway remain to be established. Here we sought to examine this question in established pancreatic cancer cell lines using the γ-secretase inhibitor IX (GSI IX) to inactivate Notch. Based on the known roles of Notch in development and stem cell biology, we focused on effects on epithelial mesenchymal transition (EMT) and on pancreatic tumor initiating CD44+/EpCAM+ cells. We analyzed the effect of the GSI IX on growth and epithelial plasticity of human pancreatic cancer cell lines, and on the tumorigenicity of pancreatic tumor initiating CD44+/EpCAM+ cells. Notably, apoptosis was induced after GSI IX treatment and EMT markers were selectively targeted. Furthermore, under GSI IX treatment, decline in the growth of pancreatic tumor initiating CD44+/EpCAM+ cells was observed <em>in vitro</em> and in a xenograft mouse model. This study demonstrates a central role of Notch signalling pathway in pancreatic cancer pathogenesis and identifies an effective approach to inhibit selectively EMT and suppress tumorigenesis by eliminating pancreatic tumor initiating CD44+/EpCAM+ cells.</p> </div>", "links"=>[], "tags"=>["epithelial", "mesenchymal", "pancreatic", "initiating", "cells", "are", "inhibited", "inhibitor", "ix"], "article_id"=>118178, "categories"=>["Cancer", "Chemistry", "Cell Biology"], "users"=>["Vindhya Palagani", "Mona El Khatib", "Uta Kossatz", "Przemyslaw Bozko", "Martin R. Müller", "Michael P. Manns", "Till Krech", "Nisar P. Malek", "Ruben R. Plentz"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0046514.s001", "https://dx.doi.org/10.1371/journal.pone.0046514.s002", "https://dx.doi.org/10.1371/journal.pone.0046514.s003", "https://dx.doi.org/10.1371/journal.pone.0046514.s004"], "stats"=>{"downloads"=>6, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Epithelial_Mesenchymal_Transition_and_Pancreatic_Tumor_Initiating_CD44_EpCAM_Cells_Are_Inhibited_by_Secretase_Inhibitor_IX/118178", "title"=>"Epithelial Mesenchymal Transition and Pancreatic Tumor Initiating CD44+/EpCAM+ Cells Are Inhibited by γ-Secretase Inhibitor IX", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-10-19 02:16:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/554873"], "description"=>"<p>(A) KP3 and (B) BxPC3 cells were treated with GSI (2.5 µM, 5 µM, 10 µM) and control (DMSO) for 96 hrs showed a down regulation of Hes1 protein by Western Blot analysis. GSI treatment resulted in a shift in the growth curves. The cell proliferation of (C) KP3 and (D) BxPC3 was measured by cell proliferation assay, GSI inhibited cell proliferation in a dose- and time-dependent manner. Note that these results reveal the anti-proliferative effects of GSI on human pancreatic cancer cells. Light microscopic pictures (10× magnification) were taken at 96 h to show the effect of GSI on cell proliferation of (E) KP3 and (F) BxPC3.</p>", "links"=>[], "tags"=>["ix", "inhibits", "proliferation", "pancreatic", "cancer", "lines", "regulates", "notch", "pathway", "downstream"], "article_id"=>225349, "categories"=>["Cancer", "Chemistry", "Cell Biology"], "users"=>["Vindhya Palagani", "Mona El Khatib", "Uta Kossatz", "Przemyslaw Bozko", "Martin R. Müller", "Michael P. Manns", "Till Krech", "Nisar P. Malek", "Ruben R. Plentz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0046514.g001", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_GSI_IX_inhibits_cell_proliferation_in_human_pancreatic_cancer_cell_lines_and_down_regulates_the_Notch_pathway_downstream_target_Hes1_/225349", "title"=>"GSI IX inhibits cell proliferation in human pancreatic cancer cell lines and down regulates the Notch pathway downstream target Hes1.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-10-19 01:29:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/555232"], "description"=>"<p>(A) KP3 and (B) BxPC3 cells were treated with control (DMSO) and GSI (2.5 µM, 5 µM and 10 µM) for 96 h. The expression of EMT markers: E-cadherin, N-cadherin, Slug and Vimentin were analyzed by Western blot. β-actin was used as a loading control. Both (A) Kp-3 and (B) BxPC3 showed no change in expression of epithelial marker E-cadherin but resulted in a GSI dose-independent down regulation of mesenchymal markers N-cadherin and Vimentin. We also detected a down regulation of the EMT transcriptional factor Slug after GSI treatment for both pancreatic cancer cell lines.</p>", "links"=>[], "tags"=>["epithelial", "mesenchymal", "markers", "gsi", "ix", "pancreatic"], "article_id"=>225718, "categories"=>["Cancer", "Chemistry", "Cell Biology"], "users"=>["Vindhya Palagani", "Mona El Khatib", "Uta Kossatz", "Przemyslaw Bozko", "Martin R. Müller", "Michael P. Manns", "Till Krech", "Nisar P. Malek", "Ruben R. Plentz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0046514.g004", "stats"=>{"downloads"=>2, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Change_in_the_expression_of_epithelial_and_mesenchymal_cell_markers_after_GSI_IX_treatment_in_human_pancreatic_cancer_/225718", "title"=>"Change in the expression of epithelial and mesenchymal cell markers after GSI IX treatment in human pancreatic cancer.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-10-19 01:35:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/555409"], "description"=>"<p>(A) Diagram shows time course of tumor growth in GSI and vehicle treated animals. The tumor growth volume was significantly decreased in the treated compared to the control group. P values were calculated with student's <i>t</i>-test; (‘*’ signifies P<0.05) error bars represent SD. (B) Pictures of mice taken at 5 weeks. The left mouse is treated with vehicle, the right mouse with GSI. The arrows are marking the growing xenografts. (C) Histology images of representative xenograft tumors from vehicle and GSI treated mice (20× magnification). (E) Ki67 staining of vehicle and GSI treated xenografts. (F) Quantification of Ki67 staining in the xenograft tumors, showing significant reduction in Ki67+ cells following GSI treatment (20× magnification). Differences were considered as statistically significant when the P-value was less <0.05 and non significant “n.s.” when the P-value was higer >0.05. Error bars show standard deviation. (D) Western Blot analysis showed a down regulation of the Notch downstream target Hes1 in xenograft tumors of GSI treated animals compared to control mice. (D) Protein expression of CD44, EpCAM, E-cadherin, N-cadherin and Slug were analyzed by the Western blot analysis and β-actin was used as a loading control. Note that <i>in vivo</i> treatment with GSI is sufficient to reverse the EMT-associated “cadherin-switch” in xenograft tumors.</p>", "links"=>[], "tags"=>["ix", "inhibits", "subcutaneously", "injected", "pancreatic"], "article_id"=>225895, "categories"=>["Cancer", "Chemistry", "Cell Biology"], "users"=>["Vindhya Palagani", "Mona El Khatib", "Uta Kossatz", "Przemyslaw Bozko", "Martin R. Müller", "Michael P. Manns", "Till Krech", "Nisar P. Malek", "Ruben R. Plentz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0046514.g006", "stats"=>{"downloads"=>3, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_GSI_IX_inhibits_the_growth_of_subcutaneously_injected_pancreatic_CD44_EpCAM_cells_/225895", "title"=>"GSI IX inhibits the growth of subcutaneously injected pancreatic CD44+/EpCAM+ cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-10-19 01:38:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/555121"], "description"=>"<p>KP3 and BxPC3 cell lines were treated for 48 h with control (DMSO) and GSI (2.5 µM, 5 µM, 10 µM) to investigate the effect of GSI on invasiveness of pancreatic cancer cell lines. The number of cells that invaded through the membrane was determined by light microscope (20× magnification) counterstained and invasion index was calculated as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0046514#s2\" target=\"_blank\">Material and Methods</a> and plotted in bar graphs. Both (A) KP3 and (B) BxPC3 showed significant decrease in number of invading cells by light microscope. Note the slight difference of invasion index between (C) Kp3 and (D) BxPC3 cells. P values are calculated with ANOVA analysis of variance along with Bonferroni post test. The error bar represents standard deviation. Differences were considered as statistically significant when the P-value was less <0.05 and non significant “n.s.” when the P-value was higher >0.05. The error bar represents standard deviation.</p>", "links"=>[], "tags"=>["ix", "attenuate", "pancreatic", "cancer"], "article_id"=>225607, "categories"=>["Cancer", "Chemistry", "Cell Biology"], "users"=>["Vindhya Palagani", "Mona El Khatib", "Uta Kossatz", "Przemyslaw Bozko", "Martin R. Müller", "Michael P. Manns", "Till Krech", "Nisar P. Malek", "Ruben R. Plentz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0046514.g003", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_GSI_IX_attenuate_invasion_of_human_pancreatic_cancer_cells_/225607", "title"=>"GSI IX attenuate invasion of human pancreatic cancer cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-10-19 01:33:27"}
  • {"files"=>["https://ndownloader.figshare.com/files/555306"], "description"=>"<p>(A) Table showing the expression of CD44+, EpCAM+ cells and the combination of CD44+/EpCAM+ cells in pancreatic cancer cell line KP3. (B) CD44+/EpCAM+ cells were treated with GSI (2.5 µM, 5 µM and 10 µM) and control (DMSO) for 48 h to determine the role of Notch in regulating the cell proliferation. Cell proliferation was inhibited in a dose- and time-dependent manner. Note that these results reveal the anti-proliferative effects of GSI on human pancreatic tumor CD44+/EpCAM+ initiating cells. (C) Light microscopic pictures (10× magnification) were taken at 48 h to show the effect of GSI on cell proliferation. (D) The down regulation of the Notch pathway was confirmed by Western Blot for Notch downstream target Hes1. Compared to unsorted and sorted DMSO treated cells Hes1 showed a dose-dependent down regulation after GSI treatment. (D) CD44 and EpCAM were down regulated in a dose dependent manner. The black arrow is marking the protein lane of CD44. (E) Epithelial marker E-cadherin was unaltered, but mesenchymal marker N-cadherin, Vimentin and Slug showed dose-dependent down regulation.</p>", "links"=>[], "tags"=>["sorted", "pancreatic", "initiating", "cells", "proliferation", "selectively", "inhibits"], "article_id"=>225788, "categories"=>["Cancer", "Chemistry", "Cell Biology"], "users"=>["Vindhya Palagani", "Mona El Khatib", "Uta Kossatz", "Przemyslaw Bozko", "Martin R. Müller", "Michael P. Manns", "Till Krech", "Nisar P. Malek", "Ruben R. Plentz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0046514.g005", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_GSI_treatment_in_sorted_pancreatic_tumor_initiating_CD44_EpCAM_cells_reduces_cell_proliferation_and_selectively_inhibits_EMT_/225788", "title"=>"GSI treatment in sorted pancreatic tumor initiating CD44+/EpCAM+ cells reduces cell proliferation and selectively inhibits EMT.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-10-19 01:36:28"}

PMC Usage Stats | Further Information

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Relative Metric

{"start_date"=>"2012-01-01T00:00:00Z", "end_date"=>"2012-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences", "average_usage"=>[322, 550, 671, 773, 864, 955, 1048, 1135, 1223, 1308, 1387, 1465, 1534, 1602, 1673, 1744, 1813, 1885, 1955, 2026, 2093, 2160, 2228, 2290, 2349]}, {"subject_area"=>"/Physical sciences/Mathematics", "average_usage"=>[325, 522, 627, 718, 804, 884, 969, 1052, 1131, 1207, 1277, 1346, 1415, 1478, 1542, 1605, 1663, 1723, 1776, 1839, 1895, 1955, 2008, 2066, 2123]}]}
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