Luteoloside Suppresses Proliferation and Metastasis of Hepatocellular Carcinoma Cells by Inhibition of NLRP3 Inflammasome
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{"title"=>"Luteoloside Suppresses Proliferation and Metastasis of Hepatocellular Carcinoma Cells by Inhibition of NLRP3 Inflammasome", "type"=>"journal", "authors"=>[{"first_name"=>"Shao-hua", "last_name"=>"Fan"}, {"first_name"=>"Yan-yan", "last_name"=>"Wang"}, {"first_name"=>"Jun", "last_name"=>"Lu"}, {"first_name"=>"Yuan-lin", "last_name"=>"Zheng"}, {"first_name"=>"Dong-mei", "last_name"=>"Wu"}, {"first_name"=>"Meng-qiu", "last_name"=>"Li"}, {"first_name"=>"Bin", "last_name"=>"Hu"}, {"first_name"=>"Zi-feng", "last_name"=>"Zhang"}, {"first_name"=>"Wei", "last_name"=>"Cheng"}, {"first_name"=>"Qun", "last_name"=>"Shan"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"1932-6203", "pmid"=>"24587153", "doi"=>"10.1371/journal.pone.0089961"}, "id"=>"78b987fe-f92e-314f-9c7e-2b02359f6e58", "abstract"=>"The inflammasome is a multi-protein complex which when activated regulates caspase-1 activation and IL-1β secretion. Inflammasome activation is mediated by NLR proteins that respond to stimuli. Among NLRs, NLRP3 senses the widest array of stimuli. NLRP3 inflammasome plays an important role in the development of many cancer types. However, Whether NLRP3 inflammasome plays an important role in the process of hepatocellular carcinoma (HCC) is still unknown. Here, the anticancer effect of luteoloside, a naturally occurring flavonoid isolated from the medicinal plant Gentiana macrophylla, against HCC cells and the underlying mechanisms were investigated. Luteoloside significantly inhibited the proliferation of HCC cells in vitro and in vivo. Live-cell imaging and transwell assays showed that the migration and invasive capacities of HCC cells, which were treated with luteoloside, were significantly inhibited compared with the control cells. The inhibitory effect of luteoloside on metastasis was also observed in vivo in male BALB/c-nu/nu mouse lung metastasis model. Further studies showed that luteoloside could significantly reduce the intracellular reactive oxygen species (ROS) accumulation. The decreased levels of ROS induced by luteoloside was accompanied by decrease in expression of NLRP3 inflammasome resulting in decrease in proteolytic cleavage of caspase-1. Inactivation of caspase-1 by luteoloside resulted in inhibition of IL-1β. Thus, luteoloside exerts its inhibitory effect on proliferation, invasion and metastasis of HCC cells through inhibition of NLRP3 inflammasome. Our results indicate that luteoloside can be a potential therapeutic agent not only as an adjuvant therapy for HCC, but also, in the control and prevention of metastatic HCC.", "link"=>"http://www.mendeley.com/research/luteoloside-suppresses-proliferation-metastasis-hepatocellular-carcinoma-cells-inhibition-nlrp3-infl", "reader_count"=>10, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Researcher"=>1, "Student > Ph. D. Student"=>4, "Student > Bachelor"=>2, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Researcher"=>1, "Student > Ph. D. Student"=>4, "Student > Bachelor"=>2, "Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Biochemistry, Genetics and Molecular Biology"=>1, "Agricultural and Biological Sciences"=>4, "Medicine and Dentistry"=>4}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>4}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>1}}, "reader_count_by_country"=>{"Japan"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1398939"], "description"=>"<p>(A) Subcutaneous injection of SMMC-7721 cells plus luteoloside treatment in nude mice inhibited tumor growth. (B) Tail vein injection of SMMC-7721 cells plus luteoloside treatment in nude mice inhibited the metastasis of SMMC-7721 cells. (a) 2×10<sup>6</sup> SMMC-7721 cells were subcutaneously injected into the right upper flank of each mouse. When tumors were observable, the animals were equally divided into two groups (ten per group). The first group received only 0.2 ml of vehicle material by gavage daily and served as a control group. The second group of animals received luteoloside (2 mg/kg body weight) in vehicle, respectively, for 4 weeks. At the termination of the experiment, the mice were sacrificed, and the tumors were weighed immediately after dissection. The yellow arrow shows the tumor. (b) The photo of tumors isolated from killed nude mice of the indicated groups. (c-d) The volume and weight of the tumors. (e) Number of metastatic nodules on the surface of the lungs of mice injected with SMMC-7721 (n = 10 mice in per group) are presented as the means and SEM. (f-g) Representative pictures of lungs with or without metastatic nodules are shown (H&E staining). *, <i>P</i><0.05; **, <i>P</i><0.01; ***, <i>P</i><0.001, versus non-luteoloside-treated control group. Scale bar: 30 µm.</p>", "links"=>[], "tags"=>["developmental biology", "morphogenesis", "cell migration", "Model organisms", "Animal models", "mouse", "Molecular cell biology", "Cell growth", "oncology", "Basic cancer research", "metastasis", "Cancers and neoplasms", "Gastrointestinal tumors", "Hepatocellular carcinoma", "inhibits", "tumorigenic", "spontaneous", "metastatic", "capabilities", "smmc-7721"], "article_id"=>944926, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Shao-hua Fan", "Yan-yan Wang", "Jun Lu", "Yuan-lin Zheng", "Dong-Mei Wu", "Meng-qiu Li", "Bin Hu", "Zi-Feng Zhang", "Wei Cheng", "Qun Shan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0089961.g005", "stats"=>{"downloads"=>0, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Luteoloside_inhibits_tumorigenic_and_spontaneous_lung_metastatic_capabilities_of_SMMC_7721_cells_/944926", "title"=>"Luteoloside inhibits tumorigenic and spontaneous lung metastatic capabilities of SMMC-7721 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-26 03:15:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/1398930"], "description"=>"<p>(A) HPLC analysis of the purity of luteoloside used in the present study. <i>Insert</i>: The chemical structure of luteoloside. (B)–(C), comparative dose- and time-dependent effect of luteoloside on the proliferation potential of HCC cells. The percentage of cell viability in different treatment groups was determined using Cell Counting Kit-8 assay. *, <i>P</i><0.05; **, <i>P</i><0.01; ***, <i>P</i><0.001; NS, not significant (<i>P</i>>0.05), versus non-luteoloside-treated control group.</p>", "links"=>[], "tags"=>["developmental biology", "morphogenesis", "cell migration", "Model organisms", "Animal models", "mouse", "Molecular cell biology", "Cell growth", "oncology", "Basic cancer research", "metastasis", "Cancers and neoplasms", "Gastrointestinal tumors", "Hepatocellular carcinoma", "inhibits", "proliferation", "hcc"], "article_id"=>944917, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Shao-hua Fan", "Yan-yan Wang", "Jun Lu", "Yuan-lin Zheng", "Dong-Mei Wu", "Meng-qiu Li", "Bin Hu", "Zi-Feng Zhang", "Wei Cheng", "Qun Shan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0089961.g001", "stats"=>{"downloads"=>0, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Luteoloside_inhibits_proliferation_of_HCC_cells_/944917", "title"=>"Luteoloside inhibits proliferation of HCC cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-26 03:15:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/1398934"], "description"=>"<p>(A) Western blot analyses of NLRP3, Caspase-1 (p10) and IL-1β protein expression in Huh-7 and SMMC-7721 cells exposed two different concentrations of luteoloside for 48 h. (B–D) Relative quantitation of NLRP3, Caspase-1 (p10) and IL-1β. (E) A hypothetical cascade pathway of NLRP3 inflammasome suppressed by luteoloside. *, <i>P</i><0.05; **, <i>P</i><0.01; ***, <i>P</i><0.001, versus non-luteoloside-treated control group.</p>", "links"=>[], "tags"=>["developmental biology", "morphogenesis", "cell migration", "Model organisms", "Animal models", "mouse", "Molecular cell biology", "Cell growth", "oncology", "Basic cancer research", "metastasis", "Cancers and neoplasms", "Gastrointestinal tumors", "Hepatocellular carcinoma", "suppresses", "nlrp3", "inflammasome"], "article_id"=>944921, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Shao-hua Fan", "Yan-yan Wang", "Jun Lu", "Yuan-lin Zheng", "Dong-Mei Wu", "Meng-qiu Li", "Bin Hu", "Zi-Feng Zhang", "Wei Cheng", "Qun Shan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0089961.g004", "stats"=>{"downloads"=>1, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Luteoloside_suppresses_the_NLRP3_inflammasome_activation_/944921", "title"=>"Luteoloside suppresses the NLRP3 inflammasome activation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-26 03:15:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/1398932"], "description"=>"<p>The migration of cells into the wound was monitored in multiple wells using a CellVoyager CV1000 confocal scanner system. The images were acquired every 0.5 hour for 48 hours (Huh-7 cells) or every hour for 72 hours (SMMC-7721 cells) (see Supplemental Movies, 1–4). The images shown represent 0 hour (A, B, E, F), 48 hours (C, D) and 72 hours (G, H). The distance between the two edges of the scratch in the luteoloside-treated cells (D or H) was greater than that of the control (C or G). (I–N) Transwell migration and invasion assays of Huh-7 cells. For the transwell migration assay, 5×10<sup>4</sup> cells were placed on the top chamber of each insert with the noncoated membrane. For the invasion assay, 1×10<sup>5</sup> cells were placed on the upper chamber of each insert coated with 150 µg Matrigel (BD Biosciences, MA). Cells in both assays were trypsinized and resuspended in DMEM, and 700–900 µL of medium supplemented with 10% fetal bovine serum was injected into the lower chambers. Representative images are shown on the left (I, J, K, L), and the quantification of five randomly selected fields is shown on the right (M, N). The values shown are expressed as the mean ± SEM. **, <i>P</i><0.01, versus non-luteoloside-treated control group. Scale bar: 100 µm.</p>", "links"=>[], "tags"=>["developmental biology", "morphogenesis", "cell migration", "Model organisms", "Animal models", "mouse", "Molecular cell biology", "Cell growth", "oncology", "Basic cancer research", "metastasis", "Cancers and neoplasms", "Gastrointestinal tumors", "Hepatocellular carcinoma", "inhibits", "hcc"], "article_id"=>944919, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Shao-hua Fan", "Yan-yan Wang", "Jun Lu", "Yuan-lin Zheng", "Dong-Mei Wu", "Meng-qiu Li", "Bin Hu", "Zi-Feng Zhang", "Wei Cheng", "Qun Shan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0089961.g002", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Luteoloside_inhibits_migration_and_invasion_of_HCC_cells_/944919", "title"=>"Luteoloside inhibits migration and invasion of HCC cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-26 03:15:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/1398933"], "description"=>"<p>ROS levels were measured using the ROS assay with DCFH-DA fluorescence dye. (A–B) Cells were treated with luteoloside at the indicated concentration for 8 h, then reacted with 10 µM DCFH-DA for 20 min. DCF fluorescence was determined on a Synergy H4 microplate reader. Cells were incubated with NAC (C), H<sub>2</sub>O<sub>2</sub> (D), diamide (E) or BSO (F) for 4 h, followed by 50 µM luteoloside for 4 h. DCF was determined on a microplate reader. (G) DCFH-DA fluorescence (green) imaging of ROS in Huh-7 cells. Scale bar: 25 µm.</p>", "links"=>[], "tags"=>["developmental biology", "morphogenesis", "cell migration", "Model organisms", "Animal models", "mouse", "Molecular cell biology", "Cell growth", "oncology", "Basic cancer research", "metastasis", "Cancers and neoplasms", "Gastrointestinal tumors", "Hepatocellular carcinoma", "decreases", "intracellular"], "article_id"=>944920, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Shao-hua Fan", "Yan-yan Wang", "Jun Lu", "Yuan-lin Zheng", "Dong-Mei Wu", "Meng-qiu Li", "Bin Hu", "Zi-Feng Zhang", "Wei Cheng", "Qun Shan"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0089961.g003", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Luteoloside_decreases_intracellular_ROS_/944920", "title"=>"Luteoloside decreases intracellular ROS.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-02-26 03:15:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/1398942", "https://ndownloader.figshare.com/files/1398943", "https://ndownloader.figshare.com/files/1398944", "https://ndownloader.figshare.com/files/1398945", "https://ndownloader.figshare.com/files/1398946", "https://ndownloader.figshare.com/files/1398947"], "description"=>"<div><p>The inflammasome is a multi-protein complex which when activated regulates caspase-1 activation and IL-1β secretion. Inflammasome activation is mediated by NLR proteins that respond to stimuli. Among NLRs, NLRP3 senses the widest array of stimuli. NLRP3 inflammasome plays an important role in the development of many cancer types. However, Whether NLRP3 inflammasome plays an important role in the process of hepatocellular carcinoma (HCC) is still unknown. Here, the anticancer effect of luteoloside, a naturally occurring flavonoid isolated from the medicinal plant <i>Gentiana macrophylla</i>, against HCC cells and the underlying mechanisms were investigated. Luteoloside significantly inhibited the proliferation of HCC cells <i>in vitro</i> and <i>in vivo</i>. Live-cell imaging and transwell assays showed that the migration and invasive capacities of HCC cells, which were treated with luteoloside, were significantly inhibited compared with the control cells. The inhibitory effect of luteoloside on metastasis was also observed <i>in vivo</i> in male BALB/c-nu/nu mouse lung metastasis model. Further studies showed that luteoloside could significantly reduce the intracellular reactive oxygen species (ROS) accumulation. The decreased levels of ROS induced by luteoloside was accompanied by decrease in expression of NLRP3 inflammasome resulting in decrease in proteolytic cleavage of caspase-1. Inactivation of caspase-1 by luteoloside resulted in inhibition of IL-1β. Thus, luteoloside exerts its inhibitory effect on proliferation, invasion and metastasis of HCC cells through inhibition of NLRP3 inflammasome. Our results indicate that luteoloside can be a potential therapeutic agent not only as an adjuvant therapy for HCC, but also, in the control and prevention of metastatic HCC.</p></div>", "links"=>[], "tags"=>["developmental biology", "morphogenesis", "cell migration", "Model organisms", "Animal models", "mouse", "Molecular cell biology", "Cell growth", "oncology", "Basic cancer research", "metastasis", "Cancers and neoplasms", "Gastrointestinal tumors", "Hepatocellular carcinoma", "suppresses", "proliferation", "hepatocellular", "carcinoma", "cells", "inhibition", "nlrp3"], "article_id"=>944929, "categories"=>["Biological Sciences", "Medicine"], "users"=>["Shao-hua Fan", "Yan-yan Wang", "Jun Lu", "Yuan-lin Zheng", "Dong-Mei Wu", "Meng-qiu Li", "Bin Hu", "Zi-Feng Zhang", "Wei Cheng", "Qun Shan"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0089961.s001", "https://dx.doi.org/10.1371/journal.pone.0089961.s002", "https://dx.doi.org/10.1371/journal.pone.0089961.s003", "https://dx.doi.org/10.1371/journal.pone.0089961.s004", "https://dx.doi.org/10.1371/journal.pone.0089961.s005", "https://dx.doi.org/10.1371/journal.pone.0089961.s006"], "stats"=>{"downloads"=>0, "page_views"=>22, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Luteoloside_Suppresses_Proliferation_and_Metastasis_of_Hepatocellular_Carcinoma_Cells_by_Inhibition_of_NLRP3_Inflammasome_/944929", "title"=>"Luteoloside Suppresses Proliferation and Metastasis of Hepatocellular Carcinoma Cells by Inhibition of NLRP3 Inflammasome", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-02-26 03:15:19"}

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Relative Metric

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