A Daphnane Diterpenoid Isolated from Wikstroemia polyantha Induces an Inflammatory Response and Modulates miRNA Activity
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{"title"=>"A Daphnane Diterpenoid isolated from Wikstroemia polyantha induces an inflammatory response and modulates miRNA activity", "type"=>"journal", "authors"=>[{"first_name"=>"Anthony", "last_name"=>"Khong", "scopus_author_id"=>"35173264200"}, {"first_name"=>"Roberto", "last_name"=>"Forestieri", "scopus_author_id"=>"55222120200"}, {"first_name"=>"David E.", "last_name"=>"Williams", "scopus_author_id"=>"24523892000"}, {"first_name"=>"Brian O.", "last_name"=>"Patrick", "scopus_author_id"=>"7004871950"}, {"first_name"=>"Andrea", "last_name"=>"Olmstead", "scopus_author_id"=>"52464168400"}, {"first_name"=>"Victoria", "last_name"=>"Svinti", "scopus_author_id"=>"55260168200"}, {"first_name"=>"Emily", "last_name"=>"Schaeffer", "scopus_author_id"=>"57197970384"}, {"first_name"=>"François", "last_name"=>"Jean", "scopus_author_id"=>"7006191250"}, {"first_name"=>"Michel", "last_name"=>"Roberge", "scopus_author_id"=>"7004311802"}, {"first_name"=>"Raymond J.", "last_name"=>"Andersen", "scopus_author_id"=>"7402653800"}, {"first_name"=>"Eric", "last_name"=>"Jan", "scopus_author_id"=>"6603942994"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-84862743241", "sgr"=>"84862743241", "pui"=>"365093308", "isbn"=>"1932-6203 (Electronic)\\n1932-6203 (Linking)", "pmid"=>"22761847", "doi"=>"10.1371/journal.pone.0039621"}, "id"=>"d5dcd61c-da26-3b24-892f-bd3228c68fe0", "abstract"=>"MicroRNAs (miRNAs) are endogenously expressed single-stranded ,21–23 nucleotide RNAs that inhibit gene expression post-transcriptionally by binding imperfectly to elements usually within the 39untranslated region (39UTR) of mRNAs. Small interfering RNAs (siRNAs) mediate site-specific cleavage by binding with perfect complementarity to RNA. Here, a cell-based miRNA reporter system was developed to screen for compounds from marine and plant extracts that inhibit miRNA or siRNA activity. The daphnane diterpenoid genkwanine M (GENK) isolated from the plant Wikstroemia polyantha induces an early inflammatory response and can moderately inhibit miR-122 activity in the liver Huh-7 cell line. GENK does not alter miR-122 levels nor does it directly inhibit siRNA activity in an in vitro cleavage assay. Finally, we demonstrate that GENK can inhibit HCV infection in Huh-7 cells. In summary, the development of the cell-based miRNA sensor system should prove useful in identifying compounds that affect miRNA/siRNA activity.", "link"=>"http://www.mendeley.com/research/daphnane-diterpenoid-isolated-wikstroemia-polyantha-induces-inflammatory-response-modulates-mirna-ac", "reader_count"=>21, "reader_count_by_academic_status"=>{"Unspecified"=>4, "Professor > Associate Professor"=>1, "Researcher"=>6, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>3, "Student > Master"=>1, "Student > Bachelor"=>3, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>4, "Professor > Associate Professor"=>1, "Researcher"=>6, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>3, "Student > Master"=>1, "Student > Bachelor"=>3, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>4, "Environmental Science"=>1, "Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>9, "Medicine and Dentistry"=>2, "Chemistry"=>2, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Chemistry"=>{"Chemistry"=>2}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>9}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>4}, "Environmental Science"=>{"Environmental Science"=>1}}, "group_count"=>2}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/619115"], "description"=>"<p>(A) Northern blot analysis of reporter Renilla luciferase (RL) and RL-miR-122 mRNA levels in Huh-7 cells treated with 10 ng/mL of TNFα for the indicated times. Huh-7 cells were transiently transfected with eEF1A-RL or eEF1A-RL-122 plasmids for 24 hours before TNFα treatment. (B) Quantitation from (A) of at least three independent experiments. At 4 hours TNFα treatment, the RL-miR-122 mRNA levels were ∼1.2 fold higher than the RL mRNA levels (p-value = 0.012).</p>", "links"=>[], "tags"=>["cells"], "article_id"=>289598, "categories"=>["Molecular Biology", "Biochemistry", "Chemistry", "Virology", "Infectious Diseases"], "users"=>["Anthony Khong", "Roberto Forestieri", "David E. Williams", "Brian O. Patrick", "Andrea Olmstead", "Victoria Svinti", "Emily Schaeffer", "François Jean", "Michel Roberge", "Raymond J. Andersen", "Eric Jan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039621.g006", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Treatment_of_cells_with_TNF_945_/289598", "title"=>"Treatment of cells with TNFα.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-26 02:39:58"}
  • {"files"=>["https://ndownloader.figshare.com/files/321883", "https://ndownloader.figshare.com/files/321921", "https://ndownloader.figshare.com/files/321959", "https://ndownloader.figshare.com/files/321994", "https://ndownloader.figshare.com/files/322018", "https://ndownloader.figshare.com/files/322069", "https://ndownloader.figshare.com/files/322167", "https://ndownloader.figshare.com/files/322208"], "description"=>"<div><p>MicroRNAs (miRNAs) are endogenously expressed single-stranded ∼21–23 nucleotide RNAs that inhibit gene expression post-transcriptionally by binding imperfectly to elements usually within the 3′untranslated region (3′UTR) of mRNAs. Small interfering RNAs (siRNAs) mediate site-specific cleavage by binding with perfect complementarity to RNA. Here, a cell-based miRNA reporter system was developed to screen for compounds from marine and plant extracts that inhibit miRNA or siRNA activity. The daphnane diterpenoid genkwanine M (GENK) isolated from the plant <em>Wikstroemia polyantha</em> induces an early inflammatory response and can moderately inhibit miR-122 activity in the liver Huh-7 cell line. GENK does not alter miR-122 levels nor does it directly inhibit siRNA activity in an <em>in vitro</em> cleavage assay. Finally, we demonstrate that GENK can inhibit HCV infection in Huh-7 cells. In summary, the development of the cell-based miRNA sensor system should prove useful in identifying compounds that affect miRNA/siRNA activity.</p> </div>", "links"=>[], "tags"=>["daphnane", "diterpenoid", "induces", "inflammatory", "modulates", "mirna"], "article_id"=>123532, "categories"=>["Molecular Biology", "Cancer", "Biochemistry", "Chemistry"], "users"=>["Anthony Khong", "Roberto Forestieri", "David E. Williams", "Brian O. Patrick", "Andrea Olmstead", "Victoria Svinti", "Emily Schaeffer", "François Jean", "Michel Roberge", "Raymond J. Andersen", "Eric Jan"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0039621.s001", "https://dx.doi.org/10.1371/journal.pone.0039621.s002", "https://dx.doi.org/10.1371/journal.pone.0039621.s003", "https://dx.doi.org/10.1371/journal.pone.0039621.s004", "https://dx.doi.org/10.1371/journal.pone.0039621.s005", "https://dx.doi.org/10.1371/journal.pone.0039621.s006", "https://dx.doi.org/10.1371/journal.pone.0039621.s007", "https://dx.doi.org/10.1371/journal.pone.0039621.s008"], "stats"=>{"downloads"=>4, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/A_Daphnane_Diterpenoid_Isolated_from_Wikstroemia_polyantha_Induces_an_Inflammatory_Response_and_Modulates_miRNA_Activity/123532", "title"=>"A Daphnane Diterpenoid Isolated from <em>Wikstroemia polyantha</em> Induces an Inflammatory Response and Modulates miRNA Activity", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-06-26 00:58:52"}
  • {"files"=>["https://ndownloader.figshare.com/files/618799"], "description"=>"<p>(B) Diacetyl GENK was prepared from GENK by treatment with acetic anhydride and DMAP in anhydrous THF.</p>", "links"=>[], "tags"=>["genkwanine", "diacetyl"], "article_id"=>289286, "categories"=>["Molecular Biology", "Biochemistry", "Chemistry", "Virology", "Infectious Diseases"], "users"=>["Anthony Khong", "Roberto Forestieri", "David E. Williams", "Brian O. Patrick", "Andrea Olmstead", "Victoria Svinti", "Emily Schaeffer", "François Jean", "Michel Roberge", "Raymond J. Andersen", "Eric Jan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039621.g002", "stats"=>{"downloads"=>2, "page_views"=>72, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Structure_of_A_genkwanine_M_GENK_B_diacetyl_GENK_and_C_genkwanine_P_/289286", "title"=>"Structure of (A) genkwanine M (GENK), (B) diacetyl GENK, and (C) genkwanine P.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-26 02:34:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/619035"], "description"=>"<p>(A and B) Scatter plots of microarray-derived mRNA expression in Huh-7 cells treated with DMSO or 10 µM GENK for (A) 1 or (B) 4 hours. At the indicated times, total mRNA was isolated and analyzed as described in the Experimental Section. Normalized relative hybridization signal intensities of GENK treatment (x-axis) versus DMSO treatment (y-axis) are plotted. The data values are highly correlated (Pearson correlation 0.995 for 1 hour treatment and 0.991 for 4 hours treatment). Dotted black lines correspond to 2-fold change (up or down-regulation). (C) Venn diagram of genes up-regulated at least 2-fold after 1 and 4 hours GENK treatment. (D) Fos and Jun mRNA levels in Huh-7 cells treated with 10 µM GENK for the given times are shown by northern blot analysis. (E) Quantitation of mRNA levels from (D) normalized to GAPDH mRNA levels.</p>", "links"=>[], "tags"=>["elicited", "genk"], "article_id"=>289519, "categories"=>["Molecular Biology", "Biochemistry", "Chemistry", "Virology", "Infectious Diseases"], "users"=>["Anthony Khong", "Roberto Forestieri", "David E. Williams", "Brian O. Patrick", "Andrea Olmstead", "Victoria Svinti", "Emily Schaeffer", "François Jean", "Michel Roberge", "Raymond J. Andersen", "Eric Jan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039621.g005", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Transcriptional_response_elicited_by_GENK_treatment_/289519", "title"=>"Transcriptional response elicited by GENK treatment.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-26 02:38:39"}
  • {"files"=>["https://ndownloader.figshare.com/files/618951"], "description"=>"<p>Mature miR-122 levels (A) or let-7a miRNA levels (B) in Huh-7 and HeLa cells, respectively, treated with 10 µM GENK for the indicated times. Shown are representative Northern blots from at least three independent experiments.</p>", "links"=>[], "tags"=>["mature", "mirna"], "article_id"=>289436, "categories"=>["Molecular Biology", "Biochemistry", "Chemistry", "Virology", "Infectious Diseases"], "users"=>["Anthony Khong", "Roberto Forestieri", "David E. Williams", "Brian O. Patrick", "Andrea Olmstead", "Victoria Svinti", "Emily Schaeffer", "François Jean", "Michel Roberge", "Raymond J. Andersen", "Eric Jan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039621.g004", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_GENK_effects_on_mature_miRNA_levels_/289436", "title"=>"GENK effects on mature miRNA levels.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-26 02:37:16"}
  • {"files"=>["https://ndownloader.figshare.com/files/618869"], "description"=>"<p>(A) eGFP-let-7 mRNA levels in HeLa cells incubated with GENK for 6 hrs or transfected with AS-let-7 or AS-Ran 2′-O-methyl RNAs for 2 days. (B, C) eGFP-miR-122 mRNA levels in Huh-7 cells treated with either (B) GENK or (C) GENK P for 2 hours at the indicated concentrations (10 µM for (C)). (D, E) eGFP-miR-122 (D), eGFP (E), and GAPDH mRNA levels in Huh-7 cells treated with 10 µM of GENK for the indicated time. (F) Quantitation of eGFP-miR-122 and eGFP mRNA levels normalized to GAPDH from (D) and (E). (G) RL and RL-miR-122 reporter mRNA levels in Huh-7 cells treated with 10 µM GENK for the indicated times. (H) Quantitation of RL and RL-miR-122 mRNA levels normalized to GAPDH from (G). At 6 and 8 hours GENK treatment, the RL-miR-122 mRNA levels were ∼1.2 fold higher than the RL mRNA levels (6 hours, p-value = 0.0192; 8 hours, p-value = 0.055). For each experiment, cells were transiently transfected with the indicated reporter plasmids for 24 hours before GENK treatment. Shown are representative Northern blots from at least three independent experiments.</p>", "links"=>[], "tags"=>["rna"], "article_id"=>289354, "categories"=>["Molecular Biology", "Biochemistry", "Chemistry", "Virology", "Infectious Diseases"], "users"=>["Anthony Khong", "Roberto Forestieri", "David E. Williams", "Brian O. Patrick", "Andrea Olmstead", "Victoria Svinti", "Emily Schaeffer", "François Jean", "Michel Roberge", "Raymond J. Andersen", "Eric Jan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039621.g003", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_GENK_effects_on_reporter_RNA_levels_/289354", "title"=>"GENK effects on reporter RNA levels.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-26 02:35:54"}
  • {"files"=>["https://ndownloader.figshare.com/files/619173"], "description"=>"<p>Huh-7 cells were mock-infected or infected with HCV at a MOI of 0.25, 0.5, or 1 in the presence of DMSO or 10 µM GENK. DMSO or GENK was added 24 hrs prior to infection and subsequently every 24 hrs. 48 hrs post-infection, cells were fixed then probed with an HCV-core protein specific antibody and cell nuclei were stained with Hoechst dye. The percentage of HCV infected cells (positive for core expression) was determined by immunofluorescence using a high-throughput microscope (Cellomics). At a MOI of 0.5 and 1, 10 µM GENK treatment inhibited HCV infection as compared to DMSO treatment (p-value <0.0001). There was no difference in the percentage of cells with immunofluorescence above background in mock-infected or HCV-infected cells (MOI 0.25) treated with DMSO or 10 µM GENK (p-value = 0.2918 and p-value = 0.2484, respectively).</p>", "links"=>[], "tags"=>["hcv", "replication", "huh-7", "cells"], "article_id"=>289655, "categories"=>["Molecular Biology", "Biochemistry", "Chemistry", "Virology", "Infectious Diseases"], "users"=>["Anthony Khong", "Roberto Forestieri", "David E. Williams", "Brian O. Patrick", "Andrea Olmstead", "Victoria Svinti", "Emily Schaeffer", "François Jean", "Michel Roberge", "Raymond J. Andersen", "Eric Jan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039621.g007", "stats"=>{"downloads"=>3, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Inhibition_of_HCV_replication_in_Huh_7_cells_by_GENK_/289655", "title"=>"Inhibition of HCV replication in Huh-7 cells by GENK.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-26 02:40:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/618738"], "description"=>"<p>(A) Schematics of CMV and eEF1a promoter-driven expression plasmids that contain the eGFP or Renilla luciferase (RL) reporter gene alone or with a perfectly complementary binding site for let-7a or for miR-122 (black rectangle). (B) Stably transfected HeLa cells expressing CMV-GFP-let-7 were incubated with a series of plant or marine extracts for 8 hours or transfected with 2′ O-methyl antisense let-7 (AS-let-7) or random RNAs (AS-Ran) for 2 days. Cells were subsequently fixed and stained with Hoechst dye, and imaged by a high-throughput microscope (Cellomics). Representative images are shown of cells treated with a positive or negative extract and of cells transfected with AS-let-7 or AS-Ran.</p>", "links"=>[], "tags"=>["assay", "mirna"], "article_id"=>289220, "categories"=>["Molecular Biology", "Biochemistry", "Chemistry", "Virology", "Infectious Diseases"], "users"=>["Anthony Khong", "Roberto Forestieri", "David E. Williams", "Brian O. Patrick", "Andrea Olmstead", "Victoria Svinti", "Emily Schaeffer", "François Jean", "Michel Roberge", "Raymond J. Andersen", "Eric Jan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039621.g001", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Cell_based_assay_for_the_identification_of_small_molecule_miRNA_inhibitors_/289220", "title"=>"Cell-based assay for the identification of small molecule miRNA inhibitors.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-26 02:33:40"}

PMC Usage Stats | Further Information

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Relative Metric

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