FHL2 Silencing Reduces Wnt Signaling and Osteosarcoma Tumorigenesis In Vitro and In Vivo
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{"title"=>"FHL2 Silencing Reduces Wnt Signaling and Osteosarcoma Tumorigenesis In Vitro and In Vivo", "type"=>"journal", "authors"=>[{"first_name"=>"Julia", "last_name"=>"Brun", "scopus_author_id"=>"55309062800"}, {"first_name"=>"François Xavier", "last_name"=>"Dieudonné", "scopus_author_id"=>"36793122000"}, {"first_name"=>"Caroline", "last_name"=>"Marty", "scopus_author_id"=>"16646605800"}, {"first_name"=>"Judith", "last_name"=>"Müller", "scopus_author_id"=>"7404872657"}, {"first_name"=>"Roland", "last_name"=>"Schüle", "scopus_author_id"=>"7003893793"}, {"first_name"=>"Ana", "last_name"=>"Patiño-García", "scopus_author_id"=>"6603418527"}, {"first_name"=>"Fernando", "last_name"=>"Lecanda", "scopus_author_id"=>"6602287091"}, {"first_name"=>"Olivia", "last_name"=>"Fromigué", "scopus_author_id"=>"6603520174"}, {"first_name"=>"Pierre J.", "last_name"=>"Marie", "scopus_author_id"=>"24519072300"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84873850148", "pmid"=>"23383046", "pui"=>"368226762", "isbn"=>"1932-6203", "scopus"=>"2-s2.0-84873850148", "doi"=>"10.1371/journal.pone.0055034", "issn"=>"19326203"}, "id"=>"818bb7d4-6b57-3adf-9d0c-a1b872b71e83", "abstract"=>"BACKGROUND: The molecular mechanisms that are involved in the growth and invasiveness of osteosarcoma, an aggressive and invasive primary bone tumor, are not fully understood. The transcriptional co-factor FHL2 (four and a half LIM domains protein 2) acts as an oncoprotein or as a tumor suppressor depending on the tissue context. In this study, we investigated the role of FHL2 in tumorigenesis in osteosarcoma model.\\n\\nMETHODOLOGY/PRINCIPAL FINDINGS: Western blot analyses showed that FHL2 is expressed above normal in most human and murine osteosarcoma cells. Tissue microarray analysis revealed that FHL2 protein expression is high in human osteosarcoma and correlates with osteosarcoma aggressiveness. In murine osteosarcoma cells, FHL2 silencing using shRNA decreased canonical Wnt/β-catenin signaling and reduced the expression of Wnt responsive genes as well as of the key Wnt molecules Wnt5a and Wnt10b. This effect resulted in inhibition of osteosarcoma cell proliferation, invasion and migration in vitro. Using xenograft experiments, we showed that FHL2 silencing markedly reduced tumor growth and lung metastasis occurence in mice. The anti-oncogenic effect of FHL2 silencing in vivo was associated with reduced cell proliferation and decreased Wnt signaling in the tumors.\\n\\nCONCLUSION/SIGNIFICANCE: Our findings demonstrate that FHL2 acts as an oncogene in osteosarcoma cells and contributes to tumorigenesis through Wnt signaling. More importantly, FHL2 depletion greatly reduces tumor cell growth and metastasis, which raises the potential therapeutic interest of targeting FHL2 to efficiently impact primary bone tumors.", "link"=>"http://www.mendeley.com/research/fhl2-silencing-reduces-wnt-signaling-osteosarcoma-tumorigenesis-vitro-vivo", "reader_count"=>17, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Researcher"=>3, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>8, "Student > Postgraduate"=>1, "Student > Bachelor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Researcher"=>3, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>8, "Student > Postgraduate"=>1, "Student > Bachelor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>2, "Materials Science"=>1, "Medicine and Dentistry"=>4, "Agricultural and Biological Sciences"=>7, "Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "reader_count_by_subdiscipline"=>{"Materials Science"=>{"Materials Science"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>7}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/499895"], "description"=>"<p>Migration of shControl and shFHL2-transduced K7M2 cells was evaluated by Boyden’s chamber (A) and wounding assays (C) and migrating cell number was evaluated (B, D). K7M2 cell invasion was evaluated by the Matrigel invasion assay (E, F). *: <i>P</i><0.05 <i>vs</i> shControl-transduced cells.</p>", "links"=>[], "tags"=>["decreases"], "article_id"=>170412, "categories"=>["Cancer", "Molecular Biology", "Cell Biology"], "users"=>["Julia Brun", "François-Xavier Dieudonné", "Caroline Marty", "Judith Müller", "Roland Schüle", "Ana Patiño-García", "Fernando Lecanda", "Olivia Fromigué", "Pierre J. Marie"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055034.g004", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FHL2_silencing_decreases_bone_tumor_cell_migration_and_invasion_/170412", "title"=>"FHL2 silencing decreases bone tumor cell migration and invasion.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:22:37"}
  • {"files"=>["https://ndownloader.figshare.com/files/499686"], "description"=>"<p>Cell lysates of osteoblast precursor cell (C3H10T1/2), calvaria-derived osteoblastic cells (MC3T3E1) and osteosarcoma cell lines (K7M2) were analysed by western blot and FHL2 level was corrected for β-actin (A). After transduction with shControl or shFHL2, FHL2 levels in K7M2 cells were evaluated by q-PCR (B) and Western blot analysis (C). shControl and shFHL2 transduced K7M2 cells were treated for 24 h with Wnt3a CM and β-catenin nuclear translocation in K7M2 cells was evaluated by Western blot analysis of nuclear fraction (D), immunocytochemistry (red, arrows: β-catenin, blue: DAPI) (E), and β-catenin transcriptional activity was determined by a reporter assay (F). The mRNA levels in the shControl and shFHL2 cells were evaluated by q-PCR analysis (G, H). *: <i>P</i><0.05 <i>vs</i> the indicated group or shControl cells.</p>", "links"=>[], "tags"=>["decreases", "signaling", "osteosarcoma"], "article_id"=>170196, "categories"=>["Cancer", "Molecular Biology", "Cell Biology"], "users"=>["Julia Brun", "François-Xavier Dieudonné", "Caroline Marty", "Judith Müller", "Roland Schüle", "Ana Patiño-García", "Fernando Lecanda", "Olivia Fromigué", "Pierre J. Marie"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055034.g002", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FHL2_silencing_decreases_Wnt_946_catenin_signaling_in_osteosarcoma_cells_/170196", "title"=>"FHL2 silencing decreases Wnt/β-catenin signaling in osteosarcoma cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:21:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/500031"], "description"=>"<p>shRNA control and shFHL2-transduced murine K7M2 cells were injected in BALB/c mice and tumor size (A) and volume (B) were determined at 6 weeks (n = 9 per group). Cell proliferation and apoptosis in tumors was determined by histological analysis using Ki67 (C, D) and TUNEL staining (arrows), respectively (E, F). Wnt5a and Wnt10b mRNA expression was evaluated in the tumors by q-PCR analysis (G). *: <i>P</i><0.05 <i>vs</i> shControl cells.</p>", "links"=>[], "tags"=>["decreases"], "article_id"=>170542, "categories"=>["Cancer", "Molecular Biology", "Cell Biology"], "users"=>["Julia Brun", "François-Xavier Dieudonné", "Caroline Marty", "Judith Müller", "Roland Schüle", "Ana Patiño-García", "Fernando Lecanda", "Olivia Fromigué", "Pierre J. Marie"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055034.g005", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FHL2_silencing_decreases_bone_tumor_growth_in_vivo_/170542", "title"=>"FHL2 silencing decreases bone tumor growth <i>in vivo</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:23:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/499775"], "description"=>"<p>After treatment with Wnt3a CM (A) or FGF-2 (0.50 ng/ml<b>)</b> (B) for 3 days, DNA replication was evaluated by BrdU incorporation in shControl and shFHL2-transduced K7M2 cells. Apoptosis was induced by serum deprivation and effector caspases activity was evaluated at 48 h in cells treated with or without Wnt3a CM (C). a: p<0.05 <i>vs</i> untreated, b:p<0.05 <i>vs</i> shControl. TUNEL analysis was performed in basal and serum deprivation conditions at 72 h (D). *: <i>P</i><0.05 <i>vs</i> the indicated group or shControl cells.</p>", "links"=>[], "tags"=>["decreases", "osteosarcoma"], "article_id"=>170286, "categories"=>["Cancer", "Molecular Biology", "Cell Biology"], "users"=>["Julia Brun", "François-Xavier Dieudonné", "Caroline Marty", "Judith Müller", "Roland Schüle", "Ana Patiño-García", "Fernando Lecanda", "Olivia Fromigué", "Pierre J. Marie"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055034.g003", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FHL2_silencing_decreases_osteosarcoma_cell_growth_/170286", "title"=>"FHL2 silencing decreases osteosarcoma cell growth.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:22:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/500133"], "description"=>"<p>Histological hematoxylin/eosin (H&E) staining of lung tissue sections showing metastasis (stars) developed in mice injected IM with shControl or shFHL2-transduced K7M2 cells (A). Metastasis area (B) and number (C) in the lung tissue were evaluated. Results are expressed as mean ± s.d. (n = 9 animals per group). *<i>P</i><0.05 <i>vs</i> shControl. Proposed model in which FHL2 silencing using shFHL2 in murine osteosarcoma cells attenuates Wnt/β-catenin signaling and reduces the expression of Wnt5a and Wnt10b and possibly other FHL2 target genes in the tumors, resulting in decreased osteosarcoma cell growth, invasiveness and tumorigenesis <i>in vivo</i> (D).</p>", "links"=>[], "tags"=>["metastasis"], "article_id"=>170647, "categories"=>["Cancer", "Molecular Biology", "Cell Biology"], "users"=>["Julia Brun", "François-Xavier Dieudonné", "Caroline Marty", "Judith Müller", "Roland Schüle", "Ana Patiño-García", "Fernando Lecanda", "Olivia Fromigué", "Pierre J. Marie"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055034.g006", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FHL2_silencing_reduces_lung_metastasis_in_mice_/170647", "title"=>"FHL2 silencing reduces lung metastasis in mice.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:23:56"}
  • {"files"=>["https://ndownloader.figshare.com/files/499585"], "description"=>"<p>Whole cell lysates were probed with the indicated antibody and revealed by Western blot analysis (A). FHL2 expression was determined by immunohistochemistry in tissue sections of normal bone, primary tumors, metastatic or recurrent osteosarcoma (Mag.×125) (B). Semi-quantitative scoring of immunohistochemical staining with anti-FHL2 antibody in normal bone and osteosarcoma samples according to patient outcome (primary tumor, metastatic or recurrent osteosarcoma) (C). *<i>P</i><0.05.</p>", "links"=>[], "tags"=>["fhl2", "osteosarcoma", "cells", "microarrays"], "article_id"=>170079, "categories"=>["Cancer", "Molecular Biology", "Cell Biology"], "users"=>["Julia Brun", "François-Xavier Dieudonné", "Caroline Marty", "Judith Müller", "Roland Schüle", "Ana Patiño-García", "Fernando Lecanda", "Olivia Fromigué", "Pierre J. Marie"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055034.g001", "stats"=>{"downloads"=>0, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Basal_FHL2_expression_in_human_osteosarcoma_cells_and_in_tissue_microarrays_TMA_of_human_osteosarcomas_/170079", "title"=>"Basal FHL2 expression in human osteosarcoma cells and in tissue microarrays (TMA) of human osteosarcomas.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:20:50"}

PMC Usage Stats | Further Information

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  • {"unique-ip"=>"4", "full-text"=>"5", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"12"}
  • {"unique-ip"=>"8", "full-text"=>"8", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"11"}
  • {"unique-ip"=>"1", "full-text"=>"0", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"2"}
  • {"unique-ip"=>"6", "full-text"=>"5", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"3"}
  • {"unique-ip"=>"6", "full-text"=>"5", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"4"}
  • {"unique-ip"=>"4", "full-text"=>"6", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"5"}
  • {"unique-ip"=>"9", "full-text"=>"11", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"8"}
  • {"unique-ip"=>"47", "full-text"=>"83", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"18", "supp-data"=>"0", "cited-by"=>"5", "year"=>"2019", "month"=>"9"}
  • {"unique-ip"=>"14", "full-text"=>"5", "pdf"=>"6", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"10"}

Relative Metric

{"start_date"=>"2013-01-01T00:00:00Z", "end_date"=>"2013-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences", "average_usage"=>[269, 466, 588, 697, 800, 896, 988, 1076, 1165, 1254, 1340, 1417]}, {"subject_area"=>"/Biology and life sciences/Cell biology", "average_usage"=>[272, 472, 600, 713, 815, 911, 1004, 1094, 1185, 1273, 1358, 1441]}, {"subject_area"=>"/Biology and life sciences/Developmental biology", "average_usage"=>[275, 472, 605, 720, 822, 921, 1013, 1106, 1200, 1289, 1378, 1459, 1531]}]}
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