Resveratrol Differentially Regulates NAMPT and SIRT1 in Hepatocarcinoma Cells and Primary Human Hepatocytes
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{"title"=>"Resveratrol differentially regulates NAMPT and SIRT1 in hepatocarcinoma cells and primary human hepatocytes", "type"=>"journal", "authors"=>[{"first_name"=>"Susanne", "last_name"=>"Schuster", "scopus_author_id"=>"38661726300"}, {"first_name"=>"Melanie", "last_name"=>"Penke", "scopus_author_id"=>"56089631300"}, {"first_name"=>"Theresa", "last_name"=>"Gorski", "scopus_author_id"=>"55559560400"}, {"first_name"=>"Stefanie", "last_name"=>"Petzold-Quinque", "scopus_author_id"=>"47861414400"}, {"first_name"=>"Georg", "last_name"=>"Damm", "scopus_author_id"=>"55323035900"}, {"first_name"=>"Rolf", "last_name"=>"Gebhardt", "scopus_author_id"=>"35473401100"}, {"first_name"=>"Wieland", "last_name"=>"Kiess", "scopus_author_id"=>"7102422024"}, {"first_name"=>"Antje", "last_name"=>"Garten", "scopus_author_id"=>"12646910000"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"372725633", "issn"=>"19326203", "isbn"=>"1932-6203", "doi"=>"10.1371/journal.pone.0091045", "scopus"=>"2-s2.0-84897137060", "pmid"=>"24603648", "sgr"=>"84897137060"}, "id"=>"cb9d52f1-1755-3c6d-8ae3-51cf4e5ff1c4", "abstract"=>"Resveratrol is reported to possess chemotherapeutic properties in several cancers. In this study, we wanted to investigate the molecular mechanisms of resveratrol-induced cell cycle arrest and apoptosis as well as the impact of resveratrol on NAMPT and SIRT1 protein function and asked whether there are differences in hepatocarcinoma cells (HepG2, Hep3B cells) and non-cancerous primary human hepatocytes. We found a lower basal NAMPT mRNA and protein expression in hepatocarcinoma cells compared to primary hepatocytes. In contrast, SIRT1 was significantly higher expressed in hepatocarcinoma cells than in primary hepatocytes. Resveratrol induced cell cycle arrest in the S- and G2/M- phase and apoptosis was mediated by activation of p53 and caspase-3 in HepG2 cells. In contrast to primary hepatocytes, resveratrol treated HepG2 cells showed a reduction of NAMPT enzymatic activity and increased p53 acetylation (K382). Resveratrol induced NAMPT release from HepG2 cells which was associated with increased NAMPT mRNA expression. This effect was absent in primary hepatocytes where resveratrol was shown to function as NAMPT and SIRT1 activator. SIRT1 inhibition by EX527 resembled resveratrol effects on HepG2 cells. Furthermore, a SIRT1 overexpression significantly decreased both p53 hyperacetylation and resveratrol-induced NAMPT release as well as S-phase arrest in HepG2 cells. We could show that NAMPT and SIRT1 are differentially regulated by resveratrol in hepatocarcinoma cells and primary hepatocytes and that resveratrol did not act as a SIRT1 activator in hepatocarcinoma cells.", "link"=>"http://www.mendeley.com/research/resveratrol-differentially-regulates-nampt-sirt1-hepatocarcinoma-cells-primary-human-hepatocytes", "reader_count"=>10, "reader_count_by_academic_status"=>{"Researcher"=>3, "Student > Ph. D. Student"=>4, "Student > Master"=>1, "Other"=>1, "Student > Bachelor"=>1}, "reader_count_by_user_role"=>{"Researcher"=>3, "Student > Ph. D. Student"=>4, "Student > Master"=>1, "Other"=>1, "Student > Bachelor"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>7}, "reader_count_by_subdiscipline"=>{"Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>7}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1410005"], "description"=>"<p>A) mRNA expression and B) protein expression of NAMPT and SIRT1 in primary human hepatocytes (n = 7), HepG2 cells (n = 8) and Hep3B cells (n = 3). Representative Western Blot is shown out of three independent experiments. Measurement of C) intracellular NAD levels (left panel, primary hepatocytes n = 4, HepG2 cells n = 6), basal NAMPT enzymatic activity (middle panel, primary hepatocytes n = 3, HepG2 cells n = 4) and extracellular NAMPT (eNAMPT) levels (right panel, primary hepatocytes n = 3, HepG2 cells n = 6) in primary human hepatocytes and HepG2 cells. Data are shown as mean± SEM. Difference between two groups was evaluated using unpaired Student’s <i>t</i>-test (*p<0.05, **p<0.01, ***p<0.001).</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme metabolism", "Enzyme regulation", "Molecular cell biology", "Cell death", "Cellular stress responses", "Drugs and devices", "Pharmacodynamics", "oncology", "Cancer treatment", "Chemotherapy and drug treatment", "Cancer prevention", "sirt1", "hepatocarcinoma", "cells"], "article_id"=>953918, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Susanne Schuster", "Melanie Penke", "Theresa Gorski", "Stefanie Petzold-Quinque", "Georg Damm", "Rolf Gebhardt", "Wieland Kiess", "Antje Garten"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0091045.g001", "stats"=>{"downloads"=>0, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_NAMPT_and_SIRT1_expression_in_hepatocarcinoma_cells_and_primary_human_hepatocytes_/953918", "title"=>"NAMPT and SIRT1 expression in hepatocarcinoma cells and primary human hepatocytes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-06 03:38:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1410019"], "description"=>"<p><i>NAMPT (</i>nicotinamide phosphoribosyltransferase, also known as PBEF, visfatin); <i>p21</i>; housekeeping genes <i>beta-ACTIN</i>, <i>TBP</i> (TATA-box-binding protein) and <i>HPRT</i> (hypoxanthine phophoribosyltransferase).</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme metabolism", "Enzyme regulation", "Molecular cell biology", "Cell death", "Cellular stress responses", "Drugs and devices", "Pharmacodynamics", "oncology", "Cancer treatment", "Chemotherapy and drug treatment", "Cancer prevention", "primer", "probes", "pcr"], "article_id"=>953932, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Susanne Schuster", "Melanie Penke", "Theresa Gorski", "Stefanie Petzold-Quinque", "Georg Damm", "Rolf Gebhardt", "Wieland Kiess", "Antje Garten"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0091045.t001", "stats"=>{"downloads"=>0, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Sequences_of_Primer_and_Probes_used_for_real_time_PCR_TaqMan_/953932", "title"=>"Sequences of Primer and Probes used for <i>real-time</i> PCR (TaqMan).", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2014-03-06 03:38:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1410014"], "description"=>"<p>Acetylation of p53 (K382) in A) HepG2 cells (n = 4) and B) primary human hepatocytes (n = 3) was evaluated by Western Blot. Densitometric analysis of at least three independent experiments is shown. Data are represented as mean± SEM and statistical analysis was performed using one-way ANOVA and the Bonferroni post hoc test (*p<0.05, n.s. not significant). As a downstream target of acetylated and activated p53, the expression of p21 was analysed by Western Blot. As positive control for SIRT1 inhibition, EX527+TSA was used. SIRT1 protein expression was analysed by Western Blot in C) HepG2 cells and D) primary hepatocytes and densitometric analysis was performed. GAPDH was used as loading control, respectively. One representative blot out of at least 3 independent experiments is shown.</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme metabolism", "Enzyme regulation", "Molecular cell biology", "Cell death", "Cellular stress responses", "Drugs and devices", "Pharmacodynamics", "oncology", "Cancer treatment", "Chemotherapy and drug treatment", "Cancer prevention", "differentially", "regulates", "p53", "acetylation", "sirt1", "hepg2", "cells"], "article_id"=>953927, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Susanne Schuster", "Melanie Penke", "Theresa Gorski", "Stefanie Petzold-Quinque", "Georg Damm", "Rolf Gebhardt", "Wieland Kiess", "Antje Garten"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0091045.g006", "stats"=>{"downloads"=>1, "page_views"=>95, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Resveratrol_differentially_regulates_p53_acetylation_and_SIRT1_protein_level_in_HepG2_cells_and_primary_human_hepatocytes_/953927", "title"=>"Resveratrol differentially regulates p53 acetylation and SIRT1 protein level in HepG2 cells and primary human hepatocytes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-06 03:38:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1410015"], "description"=>"<p>Cells were stimulated with resveratrol in serum-free medium for 24 h. Supernatants of resveratrol treated A) HepG2 cells (n = 7) and B) primary human hepatocytes (n = 3) were used for quantifying extracellular NAMPT protein amount using a specific eNAMPT ELISA. eNAMPT protein concentration was normalised to the total protein amount. <i>NAMPT</i> mRNA expression in resveratrol treated C) HepG2 cells (n = 5) and D) primary human hepatocytes (n = 4) was quantified by qRT-PCR and normalised to housekeeping genes. <i>NAMPT</i> gene expression was then related to its expression in serum-free control medium (0), which was set 1. Data are represented as mean± SEM and statistical analysis was performed using one-way ANOVA and the Bonferroni post hoc test (*p<0.05; ***p<0.001; n.s. not significant). E) Supernatant of resveratrol [100 µM] or serum-free medium (con) treated HepG2 cells was used to measure NAMPT enzymatic activity and extracellular NAMPT protein levels. Counts (cpm) measured by NAMPT enzyme assay were referred to densitometric data of NAMPT protein levels in the supernatant of the same sample. Data were then normalised to serum-free control medium which was set 1. Data are shown as mean± SEM. The difference between these two groups was evaluated using unpaired Student’s <i>t</i>-test (***p<0.001).</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme metabolism", "Enzyme regulation", "Molecular cell biology", "Cell death", "Cellular stress responses", "Drugs and devices", "Pharmacodynamics", "oncology", "Cancer treatment", "Chemotherapy and drug treatment", "Cancer prevention", "resveratrol", "nampt", "mrna"], "article_id"=>953928, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Susanne Schuster", "Melanie Penke", "Theresa Gorski", "Stefanie Petzold-Quinque", "Georg Damm", "Rolf Gebhardt", "Wieland Kiess", "Antje Garten"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0091045.g007", "stats"=>{"downloads"=>5, "page_views"=>28, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_resveratrol_on_NAMPT_release_and_NAMPT_mRNA_expression_/953928", "title"=>"Effects of resveratrol on NAMPT release and NAMPT mRNA expression.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-06 03:38:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1410013"], "description"=>"<p>Cells were stimulated with resveratrol or serum-free medium (con) for 24 h. For measuring NAMPT enzymatic activity in A) HepG2 cells and (n = 4) B) primary human hepatocytes (n = 3), 50 µg of protein lysate was used for the assay and incubated for 1 h. Counts (cpm) were normalised to µg total protein. Lysates from C) HepG2 cells (n = 3) and D) primary human hepatocytes (n = 3) were used to measure NAMPT protein levels by Western Blot. Determination of intracellular NAD levels in E) HepG2 cells (n = 6) and F) primary human hepatocytes (n = 4). NAD levels were normalised to total protein amount in each sample.</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme metabolism", "Enzyme regulation", "Molecular cell biology", "Cell death", "Cellular stress responses", "Drugs and devices", "Pharmacodynamics", "oncology", "Cancer treatment", "Chemotherapy and drug treatment", "Cancer prevention", "differentially", "regulates", "nampt", "nad", "hepg2", "cells"], "article_id"=>953926, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Susanne Schuster", "Melanie Penke", "Theresa Gorski", "Stefanie Petzold-Quinque", "Georg Damm", "Rolf Gebhardt", "Wieland Kiess", "Antje Garten"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0091045.g005", "stats"=>{"downloads"=>0, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Resveratrol_differentially_regulates_NAMPT_and_NAD_levels_in_HepG2_cells_and_primary_human_hepatocytes_/953926", "title"=>"Resveratrol differentially regulates NAMPT and NAD levels in HepG2 cells and primary human hepatocytes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-06 03:38:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1410010"], "description"=>"<p>Cells were treated with resveratrol or serum-free medium (con) for 24 h. Activation of p53 through phosphorylation at serine residue 15 and cleavage of caspase-3 in A) HepG2 cells, B) Hep3B cells and C) primary human hepatocytes were analysed by Western Blot. GAPDH was used as loading control. One representative blot out of at least 3 independent experiments is shown.</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme metabolism", "Enzyme regulation", "Molecular cell biology", "Cell death", "Cellular stress responses", "Drugs and devices", "Pharmacodynamics", "oncology", "Cancer treatment", "Chemotherapy and drug treatment", "Cancer prevention", "activates", "apoptotic", "mechanisms", "hepatocarcinoma"], "article_id"=>953923, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Susanne Schuster", "Melanie Penke", "Theresa Gorski", "Stefanie Petzold-Quinque", "Georg Damm", "Rolf Gebhardt", "Wieland Kiess", "Antje Garten"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0091045.g003", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Resveratrol_activates_apoptotic_mechanisms_in_hepatocarcinoma_cells_/953923", "title"=>"Resveratrol activates apoptotic mechanisms in hepatocarcinoma cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-06 03:38:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1410021", "https://ndownloader.figshare.com/files/1410022", "https://ndownloader.figshare.com/files/1410023", "https://ndownloader.figshare.com/files/1410024", "https://ndownloader.figshare.com/files/1410025", "https://ndownloader.figshare.com/files/1410026"], "description"=>"<div><p>Resveratrol is reported to possess chemotherapeutic properties in several cancers. In this study, we wanted to investigate the molecular mechanisms of resveratrol-induced cell cycle arrest and apoptosis as well as the impact of resveratrol on NAMPT and SIRT1 protein function and asked whether there are differences in hepatocarcinoma cells (HepG2, Hep3B cells) and non-cancerous primary human hepatocytes. We found a lower basal NAMPT mRNA and protein expression in hepatocarcinoma cells compared to primary hepatocytes. In contrast, SIRT1 was significantly higher expressed in hepatocarcinoma cells than in primary hepatocytes. Resveratrol induced cell cycle arrest in the S- and G2/M- phase and apoptosis was mediated by activation of p53 and caspase-3 in HepG2 cells. In contrast to primary hepatocytes, resveratrol treated HepG2 cells showed a reduction of NAMPT enzymatic activity and increased p53 acetylation (K382). Resveratrol induced NAMPT release from HepG2 cells which was associated with increased NAMPT mRNA expression. This effect was absent in primary hepatocytes where resveratrol was shown to function as NAMPT and SIRT1 activator. SIRT1 inhibition by EX527 resembled resveratrol effects on HepG2 cells. Furthermore, a SIRT1 overexpression significantly decreased both p53 hyperacetylation and resveratrol-induced NAMPT release as well as S-phase arrest in HepG2 cells. We could show that NAMPT and SIRT1 are differentially regulated by resveratrol in hepatocarcinoma cells and primary hepatocytes and that resveratrol did not act as a SIRT1 activator in hepatocarcinoma cells.</p></div>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme metabolism", "Enzyme regulation", "Molecular cell biology", "Cell death", "Cellular stress responses", "Drugs and devices", "Pharmacodynamics", "oncology", "Cancer treatment", "Chemotherapy and drug treatment", "Cancer prevention", "differentially", "regulates", "nampt", "sirt1", "hepatocarcinoma", "cells"], "article_id"=>953934, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Susanne Schuster", "Melanie Penke", "Theresa Gorski", "Stefanie Petzold-Quinque", "Georg Damm", "Rolf Gebhardt", "Wieland Kiess", "Antje Garten"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0091045.s001", "https://dx.doi.org/10.1371/journal.pone.0091045.s002", "https://dx.doi.org/10.1371/journal.pone.0091045.s003", "https://dx.doi.org/10.1371/journal.pone.0091045.s004", "https://dx.doi.org/10.1371/journal.pone.0091045.s005", "https://dx.doi.org/10.1371/journal.pone.0091045.s006"], "stats"=>{"downloads"=>3, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Resveratrol_Differentially_Regulates_NAMPT_and_SIRT1_in_Hepatocarcinoma_Cells_and_Primary_Human_Hepatocytes_/953934", "title"=>"Resveratrol Differentially Regulates NAMPT and SIRT1 in Hepatocarcinoma Cells and Primary Human Hepatocytes", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-03-06 03:38:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1410018"], "description"=>"<p>A) SIRT1 was transiently overexpressed in HepG2 cells [2.0 µg plasmid/0.5x10<sup>6</sup> cells] using the expression vector pECE_Flag-SIRT1 from addgene (plasmid 1791; <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0091045#pone.0091045-Brunet1\" target=\"_blank\">[35]</a>). Lysates of cells transfected with the empty vector pECE (mock-control) (1) or pECE Flag-SIRT1 vector (2) were used for Western Blot analysis. B) mock-transfected (mock-control) and Flag-SIRT1 transfected HepG2 cells were stimulated with resveratrol [50, 100 µM Resv.] for 24 h and Western Blot analysis of acetylated p53 (K382), p21 and GAPDH was performed. Densitometric anaylsis of acetylated p53 of three independent Western Blots is shown. Data were normalised to non-transfected HepG2 cells stimulated with resveratrol alone which was set 1. C) To analyse the effect of SIRT1 overexpression on resveratrol-induced NAMPT release, supernatant of mock-transfected and Flag-SIRT1 transfected HepG2 cells stimulated with or without resveratrol [100 µM] were used to measure eNAMPT level. One representative Western blot out of 3 independent experiments is shown. D) Cell viability of mock-transfected and Flag-SIRT1 transfected HepG2 cells treated with resveratrol [100 µM] (black bars) was measured using WST-1 assay (n = 3). Data were normalised to untreated mock-control which was set 1. E) mock-transfected (white bars) and Flag-SIRT1 transfected HepG2 cells (black bars) were stimulated with resveratrol [25, 50 µM] for 24 h. Percentage of cells in the S-phase were measured by PI staining and FACS analysis. All data are shown as mean± SEM (n = 4). The difference between two groups was evaluated using unpaired Student’s <i>t</i>-test (##p<0.01 mock-transfected cells compared to mock-transfected cells treated with resveratrol (white bars, mock-control), **p<0.01, ***p<0.001 Flag-SIRT1 transfected cells treated with resveratrol (black bars) compared to resveratrol-treated mock-transfected cells (white bars).</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme metabolism", "Enzyme regulation", "Molecular cell biology", "Cell death", "Cellular stress responses", "Drugs and devices", "Pharmacodynamics", "oncology", "Cancer treatment", "Chemotherapy and drug treatment", "Cancer prevention", "overexpression", "hepg2", "cells", "reversed", "resveratrol-induced", "sirt1", "nampt", "s-phase"], "article_id"=>953931, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Susanne Schuster", "Melanie Penke", "Theresa Gorski", "Stefanie Petzold-Quinque", "Georg Damm", "Rolf Gebhardt", "Wieland Kiess", "Antje Garten"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0091045.g009", "stats"=>{"downloads"=>0, "page_views"=>33, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_SIRT1_overexpression_in_HepG2_cells_reversed_resveratrol_induced_SIRT1_inhibition_NAMPT_release_and_S_phase_arrest_/953931", "title"=>"SIRT1 overexpression in HepG2 cells reversed resveratrol-induced SIRT1 inhibition, NAMPT release and S-phase arrest.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-06 03:38:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1410017"], "description"=>"<p>HepG2 cells were treated with EX527+TSA [20 µM EX527+1 µM TSA] or serum-free medium (con) for 24 h. Measurement of A) NAMPT enzymatic activity (n = 3). Counts (cpm) were normalised to µg total protein in each sample (*p<0.05). B) NAD level were determined by HPLC (n = 5) and normalised to total protein amount in each sample. C) Supernatant of EX527 treated HepG2 cells was used for determination of eNAMPT level. One representative Western blot out of 3 independent experiments is shown.</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme metabolism", "Enzyme regulation", "Molecular cell biology", "Cell death", "Cellular stress responses", "Drugs and devices", "Pharmacodynamics", "oncology", "Cancer treatment", "Chemotherapy and drug treatment", "Cancer prevention", "inhibition", "downregulates", "nampt", "hepg2"], "article_id"=>953930, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Susanne Schuster", "Melanie Penke", "Theresa Gorski", "Stefanie Petzold-Quinque", "Georg Damm", "Rolf Gebhardt", "Wieland Kiess", "Antje Garten"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0091045.g008", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_SIRT1_inhibition_downregulates_NAMPT_activity_and_induces_NAMPT_release_in_HepG2_cells_/953930", "title"=>"SIRT1 inhibition downregulates NAMPT activity and induces NAMPT release in HepG2 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-06 03:38:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1410011"], "description"=>"<p>Cells were stimulated with FK866 [10 nM] or EX527+TSA [20 µM EX527+1 µM TSA] in serum-free medium (con). Cells treated with A) FK866 and expression of acetylated p53 (K382) after 24 h. B) Cell viability of HepG2 cells after stimulation with FK866 for 48 h measured by WST-1 assay (n = 4). Data were normalised to serum-free medium (con) which was set 1 (**p<0.01; ***p<0.001 compared to serum free medium). C) Expression of acetylated p53 (K382), p21 protein and cleavage of caspase-3 were analysed in HepG2 cells treated with EX527+TSA for 24 h. GAPDH was used as loading control. One representative blot out of 3 independent experiments is shown.</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme metabolism", "Enzyme regulation", "Molecular cell biology", "Cell death", "Cellular stress responses", "Drugs and devices", "Pharmacodynamics", "oncology", "Cancer treatment", "Chemotherapy and drug treatment", "Cancer prevention", "fk866", "ex527", "p53", "acetylation", "viability", "hepg2"], "article_id"=>953924, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Susanne Schuster", "Melanie Penke", "Theresa Gorski", "Stefanie Petzold-Quinque", "Georg Damm", "Rolf Gebhardt", "Wieland Kiess", "Antje Garten"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0091045.g004", "stats"=>{"downloads"=>6, "page_views"=>222, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_FK866_and_EX527_on_p53_acetylation_and_cell_viability_in_HepG2_cells_/953924", "title"=>"Effects of FK866 and EX527 on p53 acetylation and cell viability in HepG2 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-06 03:38:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/1410006"], "description"=>"<p>Cell viability of A) primary human hepatocytes (n = 2), HepG2 and Hep3B cells (n = 3) after stimulation with resveratrol for 24 h. Data were normalised to serum-free medium control which was set 1. B) Cell cycle distribution of HepG2 cells treated with resveratrol for 24 h. A representative result is shown out of three independent experiments. A representative dot plot is given in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0091045#pone.0091045.s002\" target=\"_blank\">Fig. S2B</a>. C) Annexin V/PI apoptosis assay of HepG2 (n = 3) and Hep3B cells (n = 3) treated with resveratrol for 24 h. D) A representative dot plot of the Annexin/PI staining in HepG2 cells is shown including the mean percentage of An+ and double An+/PI+ cells of three independent experiments. Data are shown as mean± SEM and statistical analysis was performed using one-way ANOVA and the Bonferroni post hoc test (*p<0.05; **p<0.01 compared to serum-free medium).</p>", "links"=>[], "tags"=>["Biochemistry", "enzymes", "Enzyme metabolism", "Enzyme regulation", "Molecular cell biology", "Cell death", "Cellular stress responses", "Drugs and devices", "Pharmacodynamics", "oncology", "Cancer treatment", "Chemotherapy and drug treatment", "Cancer prevention", "proliferation", "apoptosis", "hepatocarcinoma", "cells", "absent"], "article_id"=>953919, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Susanne Schuster", "Melanie Penke", "Theresa Gorski", "Stefanie Petzold-Quinque", "Georg Damm", "Rolf Gebhardt", "Wieland Kiess", "Antje Garten"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0091045.g002", "stats"=>{"downloads"=>4, "page_views"=>58, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Resveratrol_reduces_cell_proliferation_and_induces_cell_cycle_arrest_and_apoptosis_in_hepatocarcinoma_cells_which_is_absent_in_primary_human_hepatocytes_/953919", "title"=>"Resveratrol reduces cell proliferation and induces cell cycle arrest and apoptosis in hepatocarcinoma cells which is absent in primary human hepatocytes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-03-06 03:38:58"}

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