A GNAS Mutation Found in Pancreatic Intraductal Papillary Mucinous Neoplasms Induces Drastic Alterations of Gene Expression Profiles with Upregulation of Mucin Genes
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{"title"=>"A GNAS mutation found in pancreatic intraductal papillary mucinous neoplasms induces drastic alterations of gene expression profiles with upregulation of mucin genes", "type"=>"journal", "authors"=>[{"first_name"=>"Hirotake", "last_name"=>"Komatsu", "scopus_author_id"=>"56768906000"}, {"first_name"=>"Etsuko", "last_name"=>"Tanji", "scopus_author_id"=>"25229269600"}, {"first_name"=>"Naoaki", "last_name"=>"Sakata", "scopus_author_id"=>"7103072259"}, {"first_name"=>"Takeshi", "last_name"=>"Aoki", "scopus_author_id"=>"55228926600"}, {"first_name"=>"Fuyuhiko", "last_name"=>"Motoi", "scopus_author_id"=>"6602471170"}, {"first_name"=>"Takeshi", "last_name"=>"Naitoh", "scopus_author_id"=>"7006611327"}, {"first_name"=>"Yu", "last_name"=>"Katayose", "scopus_author_id"=>"7003436271"}, {"first_name"=>"Shinichi", "last_name"=>"Egawa", "scopus_author_id"=>"35430817900"}, {"first_name"=>"Michiaki", "last_name"=>"Unno", "scopus_author_id"=>"21834821600"}, {"first_name"=>"Toru", "last_name"=>"Furukawa", "scopus_author_id"=>"35392252000"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"24498386", "sgr"=>"84896876454", "doi"=>"10.1371/journal.pone.0087875", "scopus"=>"2-s2.0-84896876454", "pui"=>"372485556", "issn"=>"19326203"}, "id"=>"cc38931a-6e95-3194-8179-f8483215d7d9", "abstract"=>"GNAS, a gene encoding G protein stimulating α subunit, is frequently mutated in intraductal papillary mucinous neoplasms (IPMNs), which are indolent and slow-growing pancreatic tumors that secrete abundant mucin. The GNAS mutation is not observed in conventional ductal adenocarcinomas of the pancreas. To determine the functional significance of the GNAS mutation in pancreatic ductal lineage cells, we examined in vitro phenotypes of cells of pancreatic ductal lineage, HPDE, PK-8, PCI-35, and MIA PaCa-2, with exogenous expression of either wild-type or mutated (R201H) GNAS. We found that exogenous GNAS upregulated intracellular cyclic adenine monophosphate (cAMP), particularly in mutated GNAS transfectants, and upregulated expression of MUC2 and MUC5AC in HPDE and PK-8 cells. By contrast, exogenous GNAS inhibited expression of mucin genes in PCI-35 and MIA PaCa-2 cells, despite upregulation of cAMP. We examined global gene expression profiles of some of the cells transfected with exogenous mutated GNAS (PK-8, PCI-35, and MIA PaCa-2), and found that PK-8 cells exhibited drastic alterations of the gene expression profile, which contrasted with modest alterations in PCI-35 and MIA PaCa-2 cells. To identify a cause of these different effects of exogenous mutated GNAS on phenotypes of the cells, we examined effects of interactions of the signaling pathways of G protein-coupled receptor (GPCR), mitogen-activated protein kinase (MAPK), and phosphatidylinositol 3-kinase (PI3K) on expression of mucin genes. The MAPK and PI3K pathways significantly influenced the expression of mucin genes. Exogenous GNAS did not promote cell growth but suppressed it in some of the cells. In conclusion, mutated GNAS found in IPMNs may extensively alter gene expression profiles, including expression of mucin genes, through the interaction with MAPK and PI3K pathways in pancreatic ductal cells; these changes may determine the characteristic phenotype of IPMN. PK-8 cells expressing exogenous mutated GNAS may be an ideal in vitro model of IPMN.", "link"=>"http://www.mendeley.com/research/gnas-mutation-found-pancreatic-intraductal-papillary-mucinous-neoplasms-induces-drastic-alterations", "reader_count"=>25, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>3, "Researcher"=>4, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>3, "Student > Master"=>1, "Other"=>3, "Student > Bachelor"=>1, "Professor"=>3}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>3, "Researcher"=>4, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>3, "Student > Master"=>1, "Other"=>3, "Student > Bachelor"=>1, "Professor"=>3}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>2, "Medicine and Dentistry"=>18, "Agricultural and Biological Sciences"=>3, "Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>18}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>3}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>1}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"United States"=>2, "Japan"=>2}, "group_count"=>1}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1373674"], "description"=>"<p>(A) Immunoblots of total lysates of cells transfected with the empty vector (Vec), wild-type <i>GNAS-V5</i> (GW), and mutated <i>GNAS-V5</i> (R201H; abbreviated as GM) with or without U0126, a potent mitogen-activated protein kinase kinase (MAP2K) inhibitor. (B) Cyclic AMP quantified using an enzyme immunoassay. U0126 treatment did not affect cAMP levels in PK-8 cells but downregulated cAMP in PCI-35 cells, except in the mutated <i>GNAS</i> transfectant. (C and D) A quantitative real-time PCR assay. (C) <i>MUC2</i> was consistently downregulated by U0126 in PK-8 and PCI-35 cells, regardless of the presence of exogenous <i>GNAS</i>. (D) <i>MUC5AC</i> was consistently downregulated in PCI-35 cells, regardless of the presence of exogenous <i>GNAS</i>, and upregulated by U0126 in PK-8 cells expressing exogenous mutated GNAS. Values obtained from independently duplicated experiments were plotted. Error bars indicate standard error. *p<0.05; **p<0.01.</p>", "links"=>[], "tags"=>["genetics", "Cancer genetics", "Gene function", "Molecular cell biology", "Signal transduction", "Signaling in cellular processes", "G-protein signaling", "gene expression", "Gastroenterology and hepatology", "pancreas", "oncology", "Basic cancer research", "Tumor physiology", "Cancers and neoplasms", "Gastrointestinal tumors", "Pancreatic cancer", "contributes", "mucin", "genes"], "article_id"=>924032, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Hirotake Komatsu", "Etsuko Tanji", "Naoaki Sakata", "Takeshi Aoki", "Fuyuhiko Motoi", "Takeshi Naitoh", "Yu Katayose", "Shinichi Egawa", "Michiaki Unno", "Toru Furukawa"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087875.g004", "stats"=>{"downloads"=>6, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_MAPK_activity_contributes_to_expression_of_mucin_genes_under_different_state_of_G_protein_activity_/924032", "title"=>"MAPK activity contributes to expression of mucin genes under different state of G protein activity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-03 03:24:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/1373672"], "description"=>"<p>Colony formation of cells transfected with the empty vector (Vec), wild-type <i>GNAS</i> (GW), or mutated <i>GNAS</i> (R201H; abbreviated as GM; panels A and B). (A) Images of cells incubated with a selective medium for 4 weeks. Mock denotes untransfected cells. (B) The mean values of total area of surviving colonies are shown as relative values compared to the control (Vec). Each transfection consisted of 3 plates, and mean data from independently triplicated (PK-8 cells) or duplicated (PCI-35 and MIA PaCa-2 cells) experiments were plotted with a range of one standard error. (C) A colorimetric proliferation assay showing cell proliferation after transient exogenous GNAS expression. Relative absorbance values (Day 3/Day 1) were plotted and are presented as mean ± SEM. (D) The cell cycle assay of cells with exogenous GNAS expression. *p<0.05, **p<0.01.</p>", "links"=>[], "tags"=>["genetics", "Cancer genetics", "Gene function", "Molecular cell biology", "Signal transduction", "Signaling in cellular processes", "G-protein signaling", "gene expression", "Gastroenterology and hepatology", "pancreas", "oncology", "Basic cancer research", "Tumor physiology", "Cancers and neoplasms", "Gastrointestinal tumors", "Pancreatic cancer", "gnas", "proliferation", "alterations"], "article_id"=>924030, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Hirotake Komatsu", "Etsuko Tanji", "Naoaki Sakata", "Takeshi Aoki", "Fuyuhiko Motoi", "Takeshi Naitoh", "Yu Katayose", "Shinichi Egawa", "Michiaki Unno", "Toru Furukawa"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087875.g002", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Exogenous_GNAS_does_not_stimulate_cell_proliferation_or_alterations_of_the_cell_cycle_/924030", "title"=>"Exogenous GNAS does not stimulate cell proliferation or alterations of the cell cycle.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-03 03:24:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/1373677", "https://ndownloader.figshare.com/files/1373678", "https://ndownloader.figshare.com/files/1373679", "https://ndownloader.figshare.com/files/1373680", "https://ndownloader.figshare.com/files/1373681", "https://ndownloader.figshare.com/files/1373682", "https://ndownloader.figshare.com/files/1373683"], "description"=>"<div><p><i>GNAS</i>, a gene encoding G protein stimulating α subunit, is frequently mutated in intraductal papillary mucinous neoplasms (IPMNs), which are indolent and slow-growing pancreatic tumors that secrete abundant mucin. The <i>GNAS</i> mutation is not observed in conventional ductal adenocarcinomas of the pancreas. To determine the functional significance of the <i>GNAS</i> mutation in pancreatic ductal lineage cells, we examined <i>in vitro</i> phenotypes of cells of pancreatic ductal lineage, HPDE, PK-8, PCI-35, and MIA PaCa-2, with exogenous expression of either wild-type or mutated (R201H) GNAS. We found that exogenous GNAS upregulated intracellular cyclic adenine monophosphate (cAMP), particularly in mutated <i>GNAS</i> transfectants, and upregulated expression of <i>MUC2</i> and <i>MUC5AC</i> in HPDE and PK-8 cells. By contrast, exogenous GNAS inhibited expression of mucin genes in PCI-35 and MIA PaCa-2 cells, despite upregulation of cAMP. We examined global gene expression profiles of some of the cells transfected with exogenous mutated <i>GNAS</i> (PK-8, PCI-35, and MIA PaCa-2), and found that PK-8 cells exhibited drastic alterations of the gene expression profile, which contrasted with modest alterations in PCI-35 and MIA PaCa-2 cells. To identify a cause of these different effects of exogenous mutated <i>GNAS</i> on phenotypes of the cells, we examined effects of interactions of the signaling pathways of G protein-coupled receptor (GPCR), mitogen-activated protein kinase (MAPK), and phosphatidylinositol 3-kinase (PI3K) on expression of mucin genes. The MAPK and PI3K pathways significantly influenced the expression of mucin genes. Exogenous GNAS did not promote cell growth but suppressed it in some of the cells. In conclusion, mutated <i>GNAS</i> found in IPMNs may extensively alter gene expression profiles, including expression of mucin genes, through the interaction with MAPK and PI3K pathways in pancreatic ductal cells; these changes may determine the characteristic phenotype of IPMN. PK-8 cells expressing exogenous mutated GNAS may be an ideal <i>in vitro</i> model of IPMN.</p></div>", "links"=>[], "tags"=>["genetics", "Cancer genetics", "Gene function", "Molecular cell biology", "Signal transduction", "Signaling in cellular processes", "G-protein signaling", "gene expression", "Gastroenterology and hepatology", "pancreas", "oncology", "Basic cancer research", "Tumor physiology", "Cancers and neoplasms", "Gastrointestinal tumors", "Pancreatic cancer", "mutation", "pancreatic", "intraductal", "papillary", "mucinous", "neoplasms", "drastic", "alterations", "profiles", "upregulation", "mucin", "genes"], "article_id"=>924035, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Hirotake Komatsu", "Etsuko Tanji", "Naoaki Sakata", "Takeshi Aoki", "Fuyuhiko Motoi", "Takeshi Naitoh", "Yu Katayose", "Shinichi Egawa", "Michiaki Unno", "Toru Furukawa"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0087875.s001", "https://dx.doi.org/10.1371/journal.pone.0087875.s002", "https://dx.doi.org/10.1371/journal.pone.0087875.s003", "https://dx.doi.org/10.1371/journal.pone.0087875.s004", "https://dx.doi.org/10.1371/journal.pone.0087875.s005", "https://dx.doi.org/10.1371/journal.pone.0087875.s006", "https://dx.doi.org/10.1371/journal.pone.0087875.s007"], "stats"=>{"downloads"=>7, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/A_GNAS_Mutation_Found_in_Pancreatic_Intraductal_Papillary_Mucinous_Neoplasms_Induces_Drastic_Alterations_of_Gene_Expression_Profiles_with_Upregulation_of_Mucin_Genes/924035", "title"=>"A <i>GNAS</i> Mutation Found in Pancreatic Intraductal Papillary Mucinous Neoplasms Induces Drastic Alterations of Gene Expression Profiles with Upregulation of Mucin Genes", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-02-03 03:24:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/1373676"], "description"=>"<p>(A) The regulation of <i>MUC2</i> expression in PK-8 cells was predominantly mediated by the GPCR pathway, with synergistic effects of the MAPK pathway and additive effects of the PI3K pathway. Minimal interactions existed among the 3 signaling pathways themselves. (B) <i>MUC2</i> expression in PCI-35 cells was regulated predominantly by the MAPK pathway and additively by the PI3K pathway, whereas the GPCR pathway was antagonistic. Active interactions existed among the signaling pathways: cAMP was upregulated by active MAPK and downregulated by active PI3K. (C) <i>MUC5AC</i> expression in PK-8 cells was regulated predominantly by the GPCR pathway, the MAPK pathway was antagonistic, and the PI3K pathway played a weak role. (D) <i>MUC5AC</i> expression in PCI-35 cells was regulated predominantly by the MAPK pathway, whereas both the PI3K and GPCR pathways were antagonistic.</p>", "links"=>[], "tags"=>["genetics", "Cancer genetics", "Gene function", "Molecular cell biology", "Signal transduction", "Signaling in cellular processes", "G-protein signaling", "gene expression", "Gastroenterology and hepatology", "pancreas", "oncology", "Basic cancer research", "Tumor physiology", "Cancers and neoplasms", "Gastrointestinal tumors", "Pancreatic cancer", "pathways", "pk-8", "pci-35"], "article_id"=>924034, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Hirotake Komatsu", "Etsuko Tanji", "Naoaki Sakata", "Takeshi Aoki", "Fuyuhiko Motoi", "Takeshi Naitoh", "Yu Katayose", "Shinichi Egawa", "Michiaki Unno", "Toru Furukawa"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087875.g006", "stats"=>{"downloads"=>1, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Diverse_effects_of_signaling_pathways_on_MUC2_and_MUC5AC_expression_in_PK_8_and_PCI_35_cells_/924034", "title"=>"Diverse effects of signaling pathways on <i>MUC2</i> and <i>MUC5AC</i> expression in PK-8 and PCI-35 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-03 03:24:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/1373671"], "description"=>"<p>(A) Immunoblots of total lysates of cells transfected with the empty vector (Vec), wild-type <i>GNAS-V5</i> (GW), or mutated <i>GNAS-V5</i> (R201H; abbreviated as GM) are shown on the right. Double bands were observed in assays with the anti-G protein α antibody, where the upper bands indicate specific immunoreactivity of G protein α (arrowheads). (B) Cyclic AMP levels were determined using an enzyme immunoassay. (C and D) Quantitative real-time PCR analysis of <i>MUC2</i> (C) and <i>MUC5AC</i> (D) expression. The data are shown on a logarithmic scale and values obtained are from independently duplicated experiments. Error bars indicate standard error. *p<0.05, **p<0.01.</p>", "links"=>[], "tags"=>["genetics", "Cancer genetics", "Gene function", "Molecular cell biology", "Signal transduction", "Signaling in cellular processes", "G-protein signaling", "gene expression", "Gastroenterology and hepatology", "pancreas", "oncology", "Basic cancer research", "Tumor physiology", "Cancers and neoplasms", "Gastrointestinal tumors", "Pancreatic cancer", "gnas", "alters", "mucin", "cells", "pancreatic", "ductal", "lines", "mia"], "article_id"=>924029, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Hirotake Komatsu", "Etsuko Tanji", "Naoaki Sakata", "Takeshi Aoki", "Fuyuhiko Motoi", "Takeshi Naitoh", "Yu Katayose", "Shinichi Egawa", "Michiaki Unno", "Toru Furukawa"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087875.g001", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Exogenous_GNAS_increases_cAMP_levels_and_alters_mucin_gene_expression_in_cells_of_pancreatic_ductal_lineage_the_cell_lines_HPDE_PK_8_PCI_35_and_MIA_PaCa_2_/924029", "title"=>"Exogenous GNAS increases cAMP levels and alters mucin gene expression in cells of pancreatic ductal lineage: the cell lines HPDE, PK-8, PCI-35, and MIA PaCa-2.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-03 03:24:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/1373675"], "description"=>"<p>(A) Immunoblots of total lysates of cells transfected with the empty vector (Vec), wild-type <i>GNAS-V5</i> (GW), and mutated <i>GNAS-V5</i> (R201H; abbreviated as GM) with or without LY294002, a specific inhibitor of PI3 kinase. (B) Cyclic AMP measured by means of an enzyme immunoassay. The cAMP production was not significantly affected by LY294002 in PK-8 cells but was upregulated in PCI-35 cells. (C and D) A quantitative real-time PCR assay. <i>MUC2</i> is modestly downregulated by LY294002. The latter downregulated <i>MUC5AC</i> in PK-8 cells but upregulated it in PCI-35 cells; the effect was not significant in the PCI-35 clone expressing exogenous GNAS. Values obtained from independently duplicated experiments were plotted. Error bars indicate standard error. *p<0.05, **p<0.01.</p>", "links"=>[], "tags"=>["genetics", "Cancer genetics", "Gene function", "Molecular cell biology", "Signal transduction", "Signaling in cellular processes", "G-protein signaling", "gene expression", "Gastroenterology and hepatology", "pancreas", "oncology", "Basic cancer research", "Tumor physiology", "Cancers and neoplasms", "Gastrointestinal tumors", "Pancreatic cancer", "influences", "mucin"], "article_id"=>924033, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Hirotake Komatsu", "Etsuko Tanji", "Naoaki Sakata", "Takeshi Aoki", "Fuyuhiko Motoi", "Takeshi Naitoh", "Yu Katayose", "Shinichi Egawa", "Michiaki Unno", "Toru Furukawa"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087875.g005", "stats"=>{"downloads"=>1, "page_views"=>35, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_PI3K_AKT_activity_influences_mucin_gene_expression_under_different_state_of_G_protein_activity_/924033", "title"=>"PI3K-AKT activity influences mucin gene expression under different state of G protein activity.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-03 03:24:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/1373673"], "description"=>"<p>Genes in the PI3K-AKT and MAPK signaling pathways in KEGG Mapper (<a href=\"http://www.genome.jp/kegg/\" target=\"_blank\">http://www.genome.jp/kegg/</a>) <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087875#pone.0087875-Kanehisa1\" target=\"_blank\">[22]</a> were mapped with label colors according to the ratio of expression in cells carrying mutated <i>GNAS</i> (GM) to that in cells transfected with the empty vector (Vec).</p>", "links"=>[], "tags"=>["genetics", "Cancer genetics", "Gene function", "Molecular cell biology", "Signal transduction", "Signaling in cellular processes", "G-protein signaling", "gene expression", "Gastroenterology and hepatology", "pancreas", "oncology", "Basic cancer research", "Tumor physiology", "Cancers and neoplasms", "Gastrointestinal tumors", "Pancreatic cancer", "pathways", "exogenous", "mutated", "gnas", "pk-8", "pci-35", "mia", "paca-2"], "article_id"=>924031, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Hirotake Komatsu", "Etsuko Tanji", "Naoaki Sakata", "Takeshi Aoki", "Fuyuhiko Motoi", "Takeshi Naitoh", "Yu Katayose", "Shinichi Egawa", "Michiaki Unno", "Toru Furukawa"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0087875.g003", "stats"=>{"downloads"=>2, "page_views"=>39, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Alterations_in_gene_expression_related_to_signaling_pathways_as_a_result_of_exogenous_mutated_GNAS_R201H_expression_in_PK_8_A_PCI_35_B_and_MIA_PaCa_2_C_/924031", "title"=>"Alterations in gene expression related to signaling pathways as a result of exogenous mutated GNAS (R201H) expression in PK-8 (A), PCI-35 (B), and MIA PaCa-2 (C).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-03 03:24:33"}

PMC Usage Stats | Further Information

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Relative Metric

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