Targeting the Wnt/β-Catenin Signaling Pathway in Liver Cancer Stem Cells and Hepatocellular Carcinoma Cell Lines with FH535
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{"title"=>"Targeting the Wnt/β-catenin signaling pathway in liver cancer stem cells and hepatocellular carcinoma cell lines with FH535", "type"=>"journal", "authors"=>[{"first_name"=>"Roberto", "last_name"=>"Gedaly", "scopus_author_id"=>"23059960900"}, {"first_name"=>"Roberto", "last_name"=>"Galuppo", "scopus_author_id"=>"55758236400"}, {"first_name"=>"Michael F.", "last_name"=>"Daily", "scopus_author_id"=>"23396128000"}, {"first_name"=>"Malay", "last_name"=>"Shah", "scopus_author_id"=>"56422623300"}, {"first_name"=>"Erin", "last_name"=>"Maynard", "scopus_author_id"=>"55216429100"}, {"first_name"=>"Changguo", "last_name"=>"Chen", "scopus_author_id"=>"7501966952"}, {"first_name"=>"Xiping", "last_name"=>"Zhang", "scopus_author_id"=>"35367473800"}, {"first_name"=>"Karyn A.", "last_name"=>"Esser", "scopus_author_id"=>"7005114608"}, {"first_name"=>"Donald A.", "last_name"=>"Cohen", "scopus_author_id"=>"7404418105"}, {"first_name"=>"B. Mark", "last_name"=>"Evers", "scopus_author_id"=>"7103239079"}, {"first_name"=>"Jieyun", "last_name"=>"Jiang", "scopus_author_id"=>"55674373200"}, {"first_name"=>"Brett T.", "last_name"=>"Spear", "scopus_author_id"=>"7006254316"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"373396753", "sgr"=>"84903289148", "pmid"=>"24940873", "scopus"=>"2-s2.0-84903289148", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "doi"=>"10.1371/journal.pone.0099272", "issn"=>"19326203"}, "id"=>"8fd60c33-353a-3024-ac8f-1f36d3fc0893", "abstract"=>"Activation of the Wnt/β-catenin pathway has been observed in at least 1/3 of hepatocellular carcinomas (HCC), and a significant number of these have mutations in the β-catenin gene. Therefore, effective inhibition of this pathway could provide a novel method to treat HCC. The purposed of this study was to determine whether FH535, which was previously shown to block the β-catenin pathway, could inhibit β-catenin activation of target genes and inhibit proliferation of Liver Cancer Stem Cells (LCSC) and HCC cell lines. Using β-catenin responsive reporter genes, our data indicates that FH535 can inhibit target gene activation by endogenous and exogenously expressed β-catenin, including the constitutively active form of β-catenin that contains a Serine37Alanine mutation. Our data also indicate that proliferation of LCSC and HCC lines is inhibited by FH535 in a dose-dependent manner, and that this correlates with a decrease in the percentage of cells in S phase. Finally, we also show that expression of two well-characterized targets of β-catenin, Cyclin D1 and Survivin, is reduced by FH535. Taken together, this data indicates that FH535 has potential therapeutic value in treatment of liver cancer. Importantly, these results suggest that this therapy may be effective at several levels by targeting both HCC and LCSC.", "link"=>"http://www.mendeley.com/research/targeting-wnt%CE%B2catenin-signaling-pathway-liver-cancer-stem-cells-hepatocellular-carcinoma-cell-lines", "reader_count"=>42, "reader_count_by_academic_status"=>{"Researcher"=>9, "Student > Ph. D. Student"=>11, "Student > Postgraduate"=>3, "Student > Master"=>7, "Other"=>6, "Student > Bachelor"=>5, "Lecturer > Senior Lecturer"=>1}, "reader_count_by_user_role"=>{"Researcher"=>9, "Student > Ph. D. Student"=>11, "Student > Postgraduate"=>3, "Student > Master"=>7, "Other"=>6, "Student > Bachelor"=>5, "Lecturer > Senior Lecturer"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>3, "Engineering"=>2, "Biochemistry, Genetics and Molecular Biology"=>9, "Agricultural and Biological Sciences"=>12, "Medicine and Dentistry"=>14, "Neuroscience"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>2}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>14}, "Neuroscience"=>{"Neuroscience"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>12}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>9}, "Unspecified"=>{"Unspecified"=>3}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"Austria"=>1, "Portugal"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1539273"], "description"=>"<p>Huh7 cells were treated with DMSO alone or increasing amounts of FH535 for 38-PAGE, and transferred for Western analysis with antibodies against Cyclin D1, Survivin, and β-actin. The top of shows the western blot image; the bottom graph shows densitometric analysis of the western data. This densitometric analysis indicated that FH535 at 5 and 10 µM inhibited Cyclin D1 protein levels 28% and 64% respectively; FH535 at 5 and 10 µM inhibited Survivin protein levels 24% and 48% respectively. The experiment was done twice with similar results.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "Signaling cascades", "Wnt signaling cascade", "Beta-catenin signaling", "Molecular cell biology", "genetics", "gene expression", "Gastroenterology and hepatology", "Liver diseases", "Hepatocellular carcinoma", "oncology", "Cancers and neoplasms", "Gastrointestinal tumors", "cyclin", "d1", "survivin", "huh7"], "article_id"=>1061196, "categories"=>["Biological Sciences"], "users"=>["Roberto Gedaly", "Roberto Galuppo", "Michael F. Daily", "Malay Shah", "Erin Maynard", "Changguo Chen", "Xiping Zhang", "Karyn A. Esser", "Donald A. Cohen", "B. Mark Evers", "Jieyun Jiang", "Brett T. Spear"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0099272.g006", "stats"=>{"downloads"=>1, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FH535_reduces_cyclin_D1_and_Survivin_protein_levels_in_Huh7_cells_/1061196", "title"=>"FH535 reduces cyclin D1 and Survivin protein levels in Huh7 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 03:58:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1539270"], "description"=>"<p><b>A</b>. Huh7 cells were cultured in DMEM +10%FBS for 24 h. The cells were washed with serum free DMEM 3 times, then cultured in DMEM +0.1% FBS for 24 h for cell synchronization. Cells were then cultured in DMEM+10% FBS along with different concentrations of FH535 for 24 h. The cells were harvested and stained with propidium iodide (PI) and analyzed by flow cytometry according to the GenScript protocol (Piscataway, NJ, USA). Treatment with FH535 increased the percentage of cells in G1 and decreased the percentage of cells in S phase. The experiment was done twice with similar results. <b>B</b>. LCSC cells were cultured in CelProgen complete LCSC culture medium for 24 h. Cells were then washed with serum free CelProgen medium 3 times and cultured in CelProgen Medium +0.1% FBS for 24 h for synchronization of the cells. The cells were then returned to CelProgen Complete Medium +10% FBS with different concentrations of FH535 for 24 h. Cell cycle was assayed as per Huh7 described above.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "Signaling cascades", "Wnt signaling cascade", "Beta-catenin signaling", "Molecular cell biology", "genetics", "gene expression", "Gastroenterology and hepatology", "Liver diseases", "Hepatocellular carcinoma", "oncology", "Cancers and neoplasms", "Gastrointestinal tumors", "alters", "progression", "huh7", "lcsc"], "article_id"=>1061193, "categories"=>["Biological Sciences"], "users"=>["Roberto Gedaly", "Roberto Galuppo", "Michael F. Daily", "Malay Shah", "Erin Maynard", "Changguo Chen", "Xiping Zhang", "Karyn A. Esser", "Donald A. Cohen", "B. Mark Evers", "Jieyun Jiang", "Brett T. Spear"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0099272.g004", "stats"=>{"downloads"=>2, "page_views"=>94, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FH535_alters_cell_cycle_progression_in_Huh7_and_LCSC_cells_/1061193", "title"=>"FH535 alters cell cycle progression in Huh7 and LCSC cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 03:58:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1539268"], "description"=>"<p>Cells were seeded in 96-well plates in 0.2 ml of media as described below for 72 hours, followed by the addition of <sup>3</sup>H-thymidine at 1 µCi/well for 4 hours. Incorporation of <sup>3</sup>H-thymidine was determined by scintillation counting. In panels A, B and D, the final concentration of DMSO in each well was 0.05%; in panel C, the final DMSO concentration in each well was 0.1%. (<b><i>A</i></b>) LCSCs were plated at 1000 cells/well in DMEM with 10% FBS along with DMSO alone or with increasing amounts of FH535. (<b><i>B</i></b>). LCSCs were plated at 5000 cells/well in DMEM with 1% FBS with DMSO alone or with increasing concentrations of FH535. (<b><i>C</i></b>). LCSCs were plated in DMEM with 10% FBS at 1000 cells/well with DMSO alone or increasing concentrations of XAV939. (<b><i>D</i></b>). Huh7, Hep3B and PLC cells were plated in DMEM with 10% FBS at 1000, 2500, and 5000 cells/well, respectively, with DMSO alone or increasing concentrations of FH535. <i>P</i> values are for all the three cell lines treated with FH535 are compared to controls. The experiment was done twice with similar results.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "Signaling cascades", "Wnt signaling cascade", "Beta-catenin signaling", "Molecular cell biology", "genetics", "gene expression", "Gastroenterology and hepatology", "Liver diseases", "Hepatocellular carcinoma", "oncology", "Cancers and neoplasms", "Gastrointestinal tumors", "inhibits", "proliferation", "lcsc", "hcc"], "article_id"=>1061191, "categories"=>["Biological Sciences"], "users"=>["Roberto Gedaly", "Roberto Galuppo", "Michael F. Daily", "Malay Shah", "Erin Maynard", "Changguo Chen", "Xiping Zhang", "Karyn A. Esser", "Donald A. Cohen", "B. Mark Evers", "Jieyun Jiang", "Brett T. Spear"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0099272.g003", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FH535_inhibits_proliferation_of_LCSC_and_HCC_cell_lines_/1061191", "title"=>"FH535 inhibits proliferation of LCSC and HCC cell lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 03:58:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1539264"], "description"=>"<p>Huh7 (Panel A) and Hep3B (Panel B) HCC cells were transfected with the luciferase reporter genes TOPFlash (left panels), which contains three TCF binding sites, or E3-pGL3 (right panels), which contains the AFP enhancer element E3 that has a highly conserved TCF site. Cells were additionally co-transfected with an expression vector that contained no insert (empty vector control, E.V.), wild-type β-catenin (β-catenin), or a constitutively active form of β-catenin (βcatS37A). Renilla luciferase was used to control for variations in transfection efficiency. Six hours after the addition of DNA, cells were treated with DMSO alone (no treatment) or increasing amounts of FH535. After 48 hours, luciferase levels were determined; firefly luciferase was normalized to renilla. In both cell lines, FH535 inhibited β-catenin-dependent activation of target genes. * <i>P</i><0.05. The experiment was done twice with similar results.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "Signaling cascades", "Wnt signaling cascade", "Beta-catenin signaling", "Molecular cell biology", "genetics", "gene expression", "Gastroenterology and hepatology", "Liver diseases", "Hepatocellular carcinoma", "oncology", "Cancers and neoplasms", "Gastrointestinal tumors", "inhibits", "transcriptional", "activation", "hcc"], "article_id"=>1061187, "categories"=>["Biological Sciences"], "users"=>["Roberto Gedaly", "Roberto Galuppo", "Michael F. Daily", "Malay Shah", "Erin Maynard", "Changguo Chen", "Xiping Zhang", "Karyn A. Esser", "Donald A. Cohen", "B. Mark Evers", "Jieyun Jiang", "Brett T. Spear"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0099272.g001", "stats"=>{"downloads"=>2, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FH535_inhibits_946_catenin_dependent_transcriptional_activation_in_HCC_cell_lines_/1061187", "title"=>"FH535 inhibits β-catenin dependent transcriptional activation in HCC cell lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 03:58:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1540021"], "description"=>"<p>Cells were seeded in 96-well plates in 0.2 ml of media as described below for 72 hours, followed by the addition of <sup>3</sup>H-thymidine at 1 µCi/well for 4 hours. Incorporation of <sup>3</sup>H-thymidine was determined by scintillation counting. In panels A, B and D, the final concentration of DMSO in each well was 0.05%; in panel C, the final DMSO concentration in each well was 0.1%. (<b><i>A</i></b>) LCSCs were plated at 1000 cells/well in DMEM with 10% FBS along with DMSO alone or with increasing amounts of FH535. (<b><i>B</i></b>). LCSCs were plated at 5000 cells/well in DMEM with 1% FBS with DMSO alone or with increasing concentrations of FH535. (<b><i>C</i></b>). LCSCs were plated in DMEM with 10% FBS at 1000 cells/well with DMSO alone or increasing concentrations of XAV939. (<b><i>D</i></b>). Huh7, Hep3B and PLC cells were plated in DMEM with 10% FBS at 1000, 2500, and 5000 cells/well, respectively, with DMSO alone or increasing concentrations of FH535. <i>P</i> values are for all the three cell lines treated with FH535 are compared to controls. The experiment was done twice with similar results.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "Signaling cascades", "Wnt signaling cascade", "Beta-catenin signaling", "Molecular cell biology", "genetics", "gene expression", "Gastroenterology and hepatology", "Liver diseases", "Hepatocellular carcinoma", "oncology", "Cancers and neoplasms", "Gastrointestinal tumors", "inhibits", "proliferation", "lcsc", "hcc"], "article_id"=>1061812, "categories"=>["Biological Sciences"], "users"=>["Roberto Gedaly", "Roberto Galuppo", "Michael F. Daily", "Malay Shah", "Erin Maynard", "Changguo Chen", "Xiping Zhang", "Karyn A. Esser", "Donald A. Cohen", "B. Mark Evers", "Jieyun Jiang", "Brett T. Spear"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0099272.g003", "stats"=>{"downloads"=>2, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FH535_inhibits_proliferation_of_LCSC_and_HCC_cell_lines_/1061812", "title"=>"FH535 inhibits proliferation of LCSC and HCC cell lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 15:02:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/1539272"], "description"=>"<p>LCSCs, Huh7 and Hep3B cells were treated with DMSO alone or increasing concentrations of FH535 for 38-time PCR for expression of Cyclin D1 (Panel A) or Survivin (Panel B). In both cases, mRNA levels were plotted relative to β<sub>2</sub>-microglobulin. The experiment was done twice with similar results.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "Signaling cascades", "Wnt signaling cascade", "Beta-catenin signaling", "Molecular cell biology", "genetics", "gene expression", "Gastroenterology and hepatology", "Liver diseases", "Hepatocellular carcinoma", "oncology", "Cancers and neoplasms", "Gastrointestinal tumors", "cyclin", "d1", "survivin", "mrna", "lcsc", "hcc"], "article_id"=>1061195, "categories"=>["Biological Sciences"], "users"=>["Roberto Gedaly", "Roberto Galuppo", "Michael F. Daily", "Malay Shah", "Erin Maynard", "Changguo Chen", "Xiping Zhang", "Karyn A. Esser", "Donald A. Cohen", "B. Mark Evers", "Jieyun Jiang", "Brett T. Spear"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0099272.g005", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FH535_reduces_cyclin_D1_and_survivin_mRNA_levels_in_LCSC_and_in_HCC_cell_lines_/1061195", "title"=>"FH535 reduces cyclin D1 and survivin mRNA levels in LCSC and in HCC cell lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 03:58:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1540029", "https://ndownloader.figshare.com/files/1540030", "https://ndownloader.figshare.com/files/1540031"], "description"=>"<div><p>Activation of the Wnt/β-catenin pathway has been observed in at least 1/3 of hepatocellular carcinomas (HCC), and a significant number of these have mutations in the β-catenin gene. Therefore, effective inhibition of this pathway could provide a novel method to treat HCC. The purposed of this study was to determine whether FH535, which was previously shown to block the β-catenin pathway, could inhibit β-catenin activation of target genes and inhibit proliferation of Liver Cancer Stem Cells (LCSC) and HCC cell lines. Using β-catenin responsive reporter genes, our data indicates that FH535 can inhibit target gene activation by endogenous and exogenously expressed β-catenin, including the constitutively active form of β-catenin that contains a Serine37Alanine mutation. Our data also indicate that proliferation of LCSC and HCC lines is inhibited by FH535 in a dose-dependent manner, and that this correlates with a decrease in the percentage of cells in S phase. Finally, we also show that expression of two well-characterized targets of β-catenin, Cyclin D1 and Survivin, is reduced by FH535. Taken together, this data indicates that FH535 has potential therapeutic value in treatment of liver cancer. Importantly, these results suggest that this therapy may be effective at several levels by targeting both HCC and LCSC.</p></div>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "Signaling cascades", "Wnt signaling cascade", "Beta-catenin signaling", "Molecular cell biology", "genetics", "gene expression", "Gastroenterology and hepatology", "Liver diseases", "Hepatocellular carcinoma", "oncology", "Cancers and neoplasms", "Gastrointestinal tumors", "pathway", "cancer", "cells", "hepatocellular", "carcinoma", "lines"], "article_id"=>1061820, "categories"=>["Biological Sciences"], "users"=>["Roberto Gedaly", "Roberto Galuppo", "Michael F. Daily", "Malay Shah", "Erin Maynard", "Changguo Chen", "Xiping Zhang", "Karyn A. Esser", "Donald A. Cohen", "B. Mark Evers", "Jieyun Jiang", "Brett T. Spear"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0099272.s002", "https://dx.doi.org/10.1371/journal.pone.0099272.s003", "https://dx.doi.org/10.1371/journal.pone.0099272.s004"], "stats"=>{"downloads"=>5, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Targeting_the_Wnt_946_Catenin_Signaling_Pathway_in_Liver_Cancer_Stem_Cells_and_Hepatocellular_Carcinoma_Cell_Lines_with_FH535_/1061820", "title"=>"Targeting the Wnt/β-Catenin Signaling Pathway in Liver Cancer Stem Cells and Hepatocellular Carcinoma Cell Lines with FH535", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-06-18 15:02:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/1540026"], "description"=>"<p>Huh7 cells were treated with DMSO alone or increasing amounts of FH535 for 38-PAGE, and transferred for Western analysis with antibodies against Cyclin D1, Survivin, and β-actin. The top of shows the western blot image; the bottom graph shows densitometric analysis of the western data. This densitometric analysis indicated that FH535 at 5 and 10 µM inhibited Cyclin D1 protein levels 28% and 64% respectively; FH535 at 5 and 10 µM inhibited Survivin protein levels 24% and 48% respectively. The experiment was done twice with similar results.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "Signaling cascades", "Wnt signaling cascade", "Beta-catenin signaling", "Molecular cell biology", "genetics", "gene expression", "Gastroenterology and hepatology", "Liver diseases", "Hepatocellular carcinoma", "oncology", "Cancers and neoplasms", "Gastrointestinal tumors", "cyclin", "d1", "survivin", "huh7"], "article_id"=>1061817, "categories"=>["Biological Sciences"], "users"=>["Roberto Gedaly", "Roberto Galuppo", "Michael F. Daily", "Malay Shah", "Erin Maynard", "Changguo Chen", "Xiping Zhang", "Karyn A. Esser", "Donald A. Cohen", "B. Mark Evers", "Jieyun Jiang", "Brett T. Spear"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0099272.g006", "stats"=>{"downloads"=>2, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FH535_reduces_cyclin_D1_and_Survivin_protein_levels_in_Huh7_cells_/1061817", "title"=>"FH535 reduces cyclin D1 and Survivin protein levels in Huh7 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 15:02:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/1540024"], "description"=>"<p>LCSCs, Huh7 and Hep3B cells were treated with DMSO alone or increasing concentrations of FH535 for 38-time PCR for expression of Cyclin D1 (Panel A) or Survivin (Panel B). In both cases, mRNA levels were plotted relative to β<sub>2</sub>-microglobulin. The experiment was done twice with similar results.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "Signaling cascades", "Wnt signaling cascade", "Beta-catenin signaling", "Molecular cell biology", "genetics", "gene expression", "Gastroenterology and hepatology", "Liver diseases", "Hepatocellular carcinoma", "oncology", "Cancers and neoplasms", "Gastrointestinal tumors", "cyclin", "d1", "survivin", "mrna", "lcsc", "hcc"], "article_id"=>1061815, "categories"=>["Biological Sciences"], "users"=>["Roberto Gedaly", "Roberto Galuppo", "Michael F. Daily", "Malay Shah", "Erin Maynard", "Changguo Chen", "Xiping Zhang", "Karyn A. Esser", "Donald A. Cohen", "B. Mark Evers", "Jieyun Jiang", "Brett T. Spear"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0099272.g005", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FH535_reduces_cyclin_D1_and_survivin_mRNA_levels_in_LCSC_and_in_HCC_cell_lines_/1061815", "title"=>"FH535 reduces cyclin D1 and survivin mRNA levels in LCSC and in HCC cell lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 15:02:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/1539285", "https://ndownloader.figshare.com/files/1539286", "https://ndownloader.figshare.com/files/1539287"], "description"=>"<div><p>Activation of the Wnt/β-catenin pathway has been observed in at least 1/3 of hepatocellular carcinomas (HCC), and a significant number of these have mutations in the β-catenin gene. Therefore, effective inhibition of this pathway could provide a novel method to treat HCC. The purposed of this study was to determine whether FH535, which was previously shown to block the β-catenin pathway, could inhibit β-catenin activation of target genes and inhibit proliferation of Liver Cancer Stem Cells (LCSC) and HCC cell lines. Using β-catenin responsive reporter genes, our data indicates that FH535 can inhibit target gene activation by endogenous and exogenously expressed β-catenin, including the constitutively active form of β-catenin that contains a Serine37Alanine mutation. Our data also indicate that proliferation of LCSC and HCC lines is inhibited by FH535 in a dose-dependent manner, and that this correlates with a decrease in the percentage of cells in S phase. Finally, we also show that expression of two well-characterized targets of β-catenin, Cyclin D1 and Survivin, is reduced by FH535. Taken together, this data indicates that FH535 has potential therapeutic value in treatment of liver cancer. Importantly, these results suggest that this therapy may be effective at several levels by targeting both HCC and LCSC.</p></div>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "Signaling cascades", "Wnt signaling cascade", "Beta-catenin signaling", "Molecular cell biology", "genetics", "gene expression", "Gastroenterology and hepatology", "Liver diseases", "Hepatocellular carcinoma", "oncology", "Cancers and neoplasms", "Gastrointestinal tumors", "pathway", "cancer", "cells", "hepatocellular", "carcinoma", "lines"], "article_id"=>1061208, "categories"=>["Biological Sciences"], "users"=>["Roberto Gedaly", "Roberto Galuppo", "Michael F. Daily", "Malay Shah", "Erin Maynard", "Changguo Chen", "Xiping Zhang", "Karyn A. Esser", "Donald A. Cohen", "B. Mark Evers", "Jieyun Jiang", "Brett T. Spear"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0099272.s002", "https://dx.doi.org/10.1371/journal.pone.0099272.s003", "https://dx.doi.org/10.1371/journal.pone.0099272.s004"], "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Targeting_the_Wnt_946_Catenin_Signaling_Pathway_in_Liver_Cancer_Stem_Cells_and_Hepatocellular_Carcinoma_Cell_Lines_with_FH535_/1061208", "title"=>"Targeting the Wnt/β-Catenin Signaling Pathway in Liver Cancer Stem Cells and Hepatocellular Carcinoma Cell Lines with FH535", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-06-18 03:58:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1539266"], "description"=>"<p>LCSC (left panel), Huh7 (middle panel) and HPLC (right panel) cells were co-transfected with TOPFlash or FOPFlash luciferase reporter genes along with renilla luciferase. After 6 hours, cells were left untreated (no treatment) or treated with LiCl alone or LiCl with increasing amounts of FH535. LiCl is a known activator of β-catenin. After an additional 36 hours, cells were harvested and luciferase levels were determined; firefly luciferase was normalized to renilla. TOPFlash activity was highly induced in all three cell populations; this activation was inhibited by FH535. The negative control FOPFlash showed minimal response to LiCl or FH535. TOPFlash inhibition by FH535 was more robust in LCSC than in either HCC cell line. * <i>P</i><0.003, # P<0.001. The experiment was done twice with similar results.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "Signaling cascades", "Wnt signaling cascade", "Beta-catenin signaling", "Molecular cell biology", "genetics", "gene expression", "Gastroenterology and hepatology", "Liver diseases", "Hepatocellular carcinoma", "oncology", "Cancers and neoplasms", "Gastrointestinal tumors", "inhibits", "topflash", "activation", "lcsc", "hcc"], "article_id"=>1061189, "categories"=>["Biological Sciences"], "users"=>["Roberto Gedaly", "Roberto Galuppo", "Michael F. Daily", "Malay Shah", "Erin Maynard", "Changguo Chen", "Xiping Zhang", "Karyn A. Esser", "Donald A. Cohen", "B. Mark Evers", "Jieyun Jiang", "Brett T. Spear"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0099272.g002", "stats"=>{"downloads"=>0, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FH535_inhibits_TOPFlash_activation_in_LCSC_and_HCC_cell_lines_/1061189", "title"=>"FH535 inhibits TOPFlash activation in LCSC and HCC cell lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 03:58:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/1540023"], "description"=>"<p><b>A</b>. Huh7 cells were cultured in DMEM +10%FBS for 24 h. The cells were washed with serum free DMEM 3 times, then cultured in DMEM +0.1% FBS for 24 h for cell synchronization. Cells were then cultured in DMEM+10% FBS along with different concentrations of FH535 for 24 h. The cells were harvested and stained with propidium iodide (PI) and analyzed by flow cytometry according to the GenScript protocol (Piscataway, NJ, USA). Treatment with FH535 increased the percentage of cells in G1 and decreased the percentage of cells in S phase. The experiment was done twice with similar results. <b>B</b>. LCSC cells were cultured in CelProgen complete LCSC culture medium for 24 h. Cells were then washed with serum free CelProgen medium 3 times and cultured in CelProgen Medium +0.1% FBS for 24 h for synchronization of the cells. The cells were then returned to CelProgen Complete Medium +10% FBS with different concentrations of FH535 for 24 h. Cell cycle was assayed as per Huh7 described above.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "Signaling cascades", "Wnt signaling cascade", "Beta-catenin signaling", "Molecular cell biology", "genetics", "gene expression", "Gastroenterology and hepatology", "Liver diseases", "Hepatocellular carcinoma", "oncology", "Cancers and neoplasms", "Gastrointestinal tumors", "alters", "progression", "huh7", "lcsc"], "article_id"=>1061814, "categories"=>["Biological Sciences"], "users"=>["Roberto Gedaly", "Roberto Galuppo", "Michael F. Daily", "Malay Shah", "Erin Maynard", "Changguo Chen", "Xiping Zhang", "Karyn A. Esser", "Donald A. Cohen", "B. Mark Evers", "Jieyun Jiang", "Brett T. Spear"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0099272.g004", "stats"=>{"downloads"=>2, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FH535_alters_cell_cycle_progression_in_Huh7_and_LCSC_cells_/1061814", "title"=>"FH535 alters cell cycle progression in Huh7 and LCSC cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 15:02:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/1540020"], "description"=>"<p>LCSC (left panel), Huh7 (middle panel) and HPLC (right panel) cells were co-transfected with TOPFlash or FOPFlash luciferase reporter genes along with renilla luciferase. After 6 hours, cells were left untreated (no treatment) or treated with LiCl alone or LiCl with increasing amounts of FH535. LiCl is a known activator of β-catenin. After an additional 36 hours, cells were harvested and luciferase levels were determined; firefly luciferase was normalized to renilla. TOPFlash activity was highly induced in all three cell populations; this activation was inhibited by FH535. The negative control FOPFlash showed minimal response to LiCl or FH535. TOPFlash inhibition by FH535 was more robust in LCSC than in either HCC cell line. * <i>P</i><0.003, # P<0.001. The experiment was done twice with similar results.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "Signaling cascades", "Wnt signaling cascade", "Beta-catenin signaling", "Molecular cell biology", "genetics", "gene expression", "Gastroenterology and hepatology", "Liver diseases", "Hepatocellular carcinoma", "oncology", "Cancers and neoplasms", "Gastrointestinal tumors", "inhibits", "topflash", "activation", "lcsc", "hcc"], "article_id"=>1061811, "categories"=>["Biological Sciences"], "users"=>["Roberto Gedaly", "Roberto Galuppo", "Michael F. Daily", "Malay Shah", "Erin Maynard", "Changguo Chen", "Xiping Zhang", "Karyn A. Esser", "Donald A. Cohen", "B. Mark Evers", "Jieyun Jiang", "Brett T. Spear"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0099272.g002", "stats"=>{"downloads"=>1, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FH535_inhibits_TOPFlash_activation_in_LCSC_and_HCC_cell_lines_/1061811", "title"=>"FH535 inhibits TOPFlash activation in LCSC and HCC cell lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 15:02:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/1540018"], "description"=>"<p>Huh7 (Panel A) and Hep3B (Panel B) HCC cells were transfected with the luciferase reporter genes TOPFlash (left panels), which contains three TCF binding sites, or E3-pGL3 (right panels), which contains the AFP enhancer element E3 that has a highly conserved TCF site. Cells were additionally co-transfected with an expression vector that contained no insert (empty vector control, E.V.), wild-type β-catenin (β-catenin), or a constitutively active form of β-catenin (βcatS37A). Renilla luciferase was used to control for variations in transfection efficiency. Six hours after the addition of DNA, cells were treated with DMSO alone (no treatment) or increasing amounts of FH535. After 48 hours, luciferase levels were determined; firefly luciferase was normalized to renilla. In both cell lines, FH535 inhibited β-catenin-dependent activation of target genes. * <i>P</i><0.05. The experiment was done twice with similar results.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "Signaling cascades", "Wnt signaling cascade", "Beta-catenin signaling", "Molecular cell biology", "genetics", "gene expression", "Gastroenterology and hepatology", "Liver diseases", "Hepatocellular carcinoma", "oncology", "Cancers and neoplasms", "Gastrointestinal tumors", "inhibits", "transcriptional", "activation", "hcc"], "article_id"=>1061810, "categories"=>["Biological Sciences"], "users"=>["Roberto Gedaly", "Roberto Galuppo", "Michael F. Daily", "Malay Shah", "Erin Maynard", "Changguo Chen", "Xiping Zhang", "Karyn A. Esser", "Donald A. Cohen", "B. Mark Evers", "Jieyun Jiang", "Brett T. Spear"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0099272.g001", "stats"=>{"downloads"=>4, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FH535_inhibits_946_catenin_dependent_transcriptional_activation_in_HCC_cell_lines_/1061810", "title"=>"FH535 inhibits β-catenin dependent transcriptional activation in HCC cell lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-06-18 15:02:02"}

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