The Orphan Nuclear Receptor LRH-1 and ERα Activate GREB1 Expression to Induce Breast Cancer Cell Proliferation
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{"title"=>"The orphan nuclear receptor LRH-1 and ERα activate GREB1 expression to induce breast cancer cell proliferation", "type"=>"journal", "authors"=>[{"first_name"=>"Ashwini L.", "last_name"=>"Chand", "scopus_author_id"=>"12238803500"}, {"first_name"=>"Dhilushi D.", "last_name"=>"Wijayakumara", "scopus_author_id"=>"54924445600"}, {"first_name"=>"Kevin C.", "last_name"=>"Knower", "scopus_author_id"=>"8780217900"}, {"first_name"=>"Kerrie A.", "last_name"=>"Herridge", "scopus_author_id"=>"36704610800"}, {"first_name"=>"Tamara L.", "last_name"=>"Howard", "scopus_author_id"=>"53463451900"}, {"first_name"=>"Kyren A.", "last_name"=>"Lazarus", "scopus_author_id"=>"37087285200"}, {"first_name"=>"Colin D.", "last_name"=>"Clyne", "scopus_author_id"=>"57189756035"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84857133294", "doi"=>"10.1371/journal.pone.0031593", "pui"=>"364271770", "issn"=>"19326203", "pmid"=>"22359603", "scopus"=>"2-s2.0-84857133294"}, "id"=>"05dea49e-8519-32f8-9ae4-dd6d4188fa57", "abstract"=>"BACKGROUND: Liver Receptor Homolog 1 (LRH-1, NR5A2) is an orphan nuclear receptor that is over-expressed in cancers in tissues such as the breast, colon and pancreas. LRH-1 plays important roles in embryonic development, steroidogenesis and cholesterol homeostasis. In tumor cells, LRH-1 induces proliferation and cell cycle progression. High LRH-1 expression is demonstrated in breast cancers, positively correlating with ERα status and aromatase activity. LRH-1 dependent cellular mechanisms in breast cancer epithelial cells are poorly defined. Hence in the present study we investigated the actions of LRH-1 in estrogen receptor α (ERα) positive breast cancer cells.\\n\\nRESULTS: The study aimed to investigate LRH-1 dependent mechanisms that promote breast cancer proliferation. We identified that LRH-1 regulated the expression of Growth Regulation by Estrogen in Breast Cancer 1 (GREB1) in MCF-7 and MDA-MB-231 cells. Over-expression of LRH-1 increased GREB1 mRNA levels while knockdown of LRH-1 reduced its expression. GREB1 is a well characterised ERα target gene, with three estrogen response elements (ERE) located on its promoter. Chromatin immunoprecipitation studies provided evidence of the co-localisation of LRH-1 and ERα at all three EREs. With electrophoretic mobility shift assays, we demonstrated direct binding of LRH-1 to EREs located on GREB1 and Trefoil Factor 1 (TFF1, pS2) promoters. LRH-1 and ERα co-operatively activated transcription of ERE luciferase reporter constructs suggesting an overlap in regulation of target genes in breast cancer cells. Over-expression of LRH-1 resulted in an increase in cell proliferation. This effect was more pronounced with estradiol treatment. In the presence of ICI 182,780, an ERα antagonist, LRH-1 still induced proliferation.\\n\\nCONCLUSIONS: We conclude that in ER-positive breast cancer cells, LRH-1 promotes cell proliferation by enhancing ERα mediated transcription of target genes such as GREB-1. Collectively these findings indicate the importance of LRH-1 in the progression of hormone-dependent breast cancer and implicate LRH-1 as a potential avenue for drug development.", "link"=>"http://www.mendeley.com/research/orphan-nuclear-receptor-lrh1-er%CE%B1-activate-greb1-expression-induce-breast-cancer-cell-proliferation", "reader_count"=>38, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>2, "Researcher"=>7, "Student > Doctoral Student"=>4, "Student > Ph. D. Student"=>11, "Student > Postgraduate"=>3, "Student > Master"=>5, "Student > Bachelor"=>4, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>2, "Researcher"=>7, "Student > Doctoral Student"=>4, "Student > Ph. D. Student"=>11, "Student > Postgraduate"=>3, "Student > Master"=>5, "Student > Bachelor"=>4, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>7, "Nursing and Health Professions"=>1, "Agricultural and Biological Sciences"=>18, "Medicine and Dentistry"=>5, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Physics and Astronomy"=>1, "Chemistry"=>4}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>5}, "Chemistry"=>{"Chemistry"=>4}, "Physics and Astronomy"=>{"Physics and Astronomy"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>18}, "Nursing and Health Professions"=>{"Nursing and Health Professions"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>7}, "Unspecified"=>{"Unspecified"=>1}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"Canada"=>1, "Turkey"=>1, "United Kingdom"=>1, "Portugal"=>1}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/681809"], "description"=>"<p>(a) EMSA showing binding of LRH-1 to the EREs present in the GREB1 promoter. Radiolabeled ERE1-GREB1, ERE2-GREB1 and ERE3-GREB1 probes were incubated with <i>in vitro</i> translated LRH-1 protein. <i>In vitro</i> translation of the empty vector was used as a negative control. Anti-LRH-1 antibody was added in addition to the probe and the LRH-1 protein to indicate specificity of protein binding. (b) EMSA showing binding of LRH-1 to the EREs present in the GREB1 and pS2 promoters. Radiolabeled LRHRE probe (containing the LRH-1 response element derived from the aromatase promoter), whole cell nuclear extracts infected with a LRH-1 viral construct were incubated with various oligonucleotides (as listed in the figure) including unlabeled LRHRE, mutated LRHRE, ERE1-GREB1, ERE2-GREB1, ERE3-GREB1 and ERE-pS2 which were added in 200 fold excess. Anti-LRH-1 antibody and IgG were also added in addition to the probe and the nuclear extract to indicate specificity of protein binding.</p>", "links"=>[], "tags"=>["binds", "ere", "sequences", "greb1", "ps2"], "article_id"=>352290, "categories"=>["Cancer", "Biochemistry", "Molecular Biology"], "users"=>["Ashwini L. Chand", "Dhilushi D. Wijayakumara", "Kevin C. Knower", "Kerrie A. Herridge", "Tamara L. Howard", "Kyren A. Lazarus", "Colin D. Clyne"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0031593.g003", "stats"=>{"downloads"=>1, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/LRH_1_binds_to_specific_ERE_sequences_of_the_GREB1_and_pS2_promoters_/352290", "title"=>"LRH-1 binds to specific ERE sequences of the GREB1 and pS2 promoters.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-16 00:38:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/348161"], "description"=>"<div><h3>Background</h3><p>Liver Receptor Homolog 1 (LRH-1, NR5A2) is an orphan nuclear receptor that is over-expressed in cancers in tissues such as the breast, colon and pancreas. LRH-1 plays important roles in embryonic development, steroidogenesis and cholesterol homeostasis. In tumor cells, LRH-1 induces proliferation and cell cycle progression. High LRH-1 expression is demonstrated in breast cancers, positively correlating with ERα status and aromatase activity. LRH-1 dependent cellular mechanisms in breast cancer epithelial cells are poorly defined. Hence in the present study we investigated the actions of LRH-1 in estrogen receptor α (ERα) positive breast cancer cells.</p> <h3>Results</h3><p>The study aimed to investigate LRH-1 dependent mechanisms that promote breast cancer proliferation. We identified that LRH-1 regulated the expression of Growth Regulation by Estrogen in Breast Cancer 1 (GREB1) in MCF-7 and MDA-MB-231 cells. Over-expression of LRH-1 increased GREB1 mRNA levels while knockdown of LRH-1 reduced its expression. GREB1 is a well characterised ERα target gene, with three estrogen response elements (ERE) located on its promoter. Chromatin immunoprecipitation studies provided evidence of the co-localisation of LRH-1 and ERα at all three EREs. With electrophoretic mobility shift assays, we demonstrated direct binding of LRH-1 to EREs located on GREB1 and Trefoil Factor 1 (TFF1, pS2) promoters. LRH-1 and ERα co-operatively activated transcription of ERE luciferase reporter constructs suggesting an overlap in regulation of target genes in breast cancer cells. Over-expression of LRH-1 resulted in an increase in cell proliferation. This effect was more pronounced with estradiol treatment. In the presence of ICI 182,780, an ERα antagonist, LRH-1 still induced proliferation.</p> <h3>Conclusions</h3><p>We conclude that in ER-positive breast cancer cells, LRH-1 promotes cell proliferation by enhancing ERα mediated transcription of target genes such as GREB-1. Collectively these findings indicate the importance of LRH-1 in the progression of hormone-dependent breast cancer and implicate LRH-1 as a potential avenue for drug development.</p> </div>", "links"=>[], "tags"=>["orphan", "receptor", "lrh-1", "activate", "greb1", "induce", "cancer", "proliferation"], "article_id"=>128837, "categories"=>["Cancer", "Biochemistry", "Molecular Biology"], "users"=>["Ashwini L. Chand", "Dhilushi D. Wijayakumara", "Kevin C. Knower", "Kerrie A. Herridge", "Tamara L. Howard", "Kyren A. Lazarus", "Colin D. Clyne"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0031593", "stats"=>{"downloads"=>6, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/The_Orphan_Nuclear_Receptor_LRH_1_and_ER_Activate_GREB1_Expression_to_Induce_Breast_Cancer_Cell_Proliferation/128837", "title"=>"The Orphan Nuclear Receptor LRH-1 and ERα Activate GREB1 Expression to Induce Breast Cancer Cell Proliferation", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2012-02-16 02:27:17"}
  • {"files"=>["https://ndownloader.figshare.com/files/681718"], "description"=>"<p>(a) Location of regulatory EREs on the distal and proximal GREB1 promoter, highlighting (bold) sequence similarity of the LRH-1 nuclear receptor half site within the ERE palindrome. (b) Chromatin immunoprecipitation (ChIP) showing occupancy of LRH-1 on the three EREs where ERα binds in the presence or absence of estradiol. Immunoprecipitation was performed with anti-LRH-1 and ERα antibodies on chromatin isolated from MCF-7 cells treated with vehicle or 10 nM 17β-estradiol for 45 mins. (c) The precipitated chromatin was analyzed by quantitative real-time PCR to demonstrate relative occupancy using the delta delta C<sub>t</sub> method. Data is normalised to 10% of input. Data is represented from 3 or more separate treatments and separate ChIP experiments. (d) Sequential ChIP demonstrating co-localisation of ERα and LRH-1 on ERE1 of the <i>GREB1</i> promoter. Figures are representative of 3 or more separate ChIP experiments.</p>", "links"=>[], "tags"=>["binds", "ere", "sites", "greb1"], "article_id"=>352200, "categories"=>["Cancer", "Biochemistry", "Molecular Biology"], "users"=>["Ashwini L. Chand", "Dhilushi D. Wijayakumara", "Kevin C. Knower", "Kerrie A. Herridge", "Tamara L. Howard", "Kyren A. Lazarus", "Colin D. Clyne"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0031593.g002", "stats"=>{"downloads"=>2, "page_views"=>19, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/LRH_1_binds_to_three_ERE_sites_within_the_GREB1_promoter_/352200", "title"=>"LRH-1 binds to three ERE sites within the GREB1 promoter.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-16 00:36:40"}
  • {"files"=>["https://ndownloader.figshare.com/files/682133"], "description"=>"<p>MDA-MB-231 cells were transfected with empty vector (C) or expression vectors for LRH-1 alone (L), ERα alone (E) or both LRH-1 and ERα (L+E). Cells were treated with vehicle or 10 nM 17β-estradiol (E2) for 16 h. Quantitation of (a) LRH-1, (b) ERα and (c) GREB1 mRNA expression. Data is presented as mean+SE, n = 3 separate experiments, ***P&lt;0.001 compared to vehicle control.</p>", "links"=>[], "tags"=>["greb1", "er", "cancer"], "article_id"=>352619, "categories"=>["Cancer", "Biochemistry", "Molecular Biology"], "users"=>["Ashwini L. Chand", "Dhilushi D. Wijayakumara", "Kevin C. Knower", "Kerrie A. Herridge", "Tamara L. Howard", "Kyren A. Lazarus", "Colin D. Clyne"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0031593.g007", "stats"=>{"downloads"=>3, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/LRH_1_regulation_of_GREB1_expression_in_ER_negative_breast_cancer_cells_/352619", "title"=>"LRH-1 regulation of GREB1 expression in ER negative breast cancer cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-16 00:43:39"}
  • {"files"=>["https://ndownloader.figshare.com/files/681978"], "description"=>"<p>Cell proliferation was measured in pcDNA alone transfected, estrogen-deprived MCF-7 cells (control) or LRH-1 over-expressing (+LRH-1) MCF-7 cells treated with vehicle, 10 nM 17β-estradiol (E2) or 10 nM 17β-estradiol and 1 nM ICI 182,780, an ERα antagonist for 5 days. Data is presented as mean+SEM, n = 3 separate experiments, triplicate treatments per experiment, ***P&lt;0.001 compared to control transfected cells; a,b P&lt;0.001 compared to vehicle control.</p>", "links"=>[], "tags"=>["induces", "proliferation", "ici", "treated"], "article_id"=>352462, "categories"=>["Cancer", "Biochemistry", "Molecular Biology"], "users"=>["Ashwini L. Chand", "Dhilushi D. Wijayakumara", "Kevin C. Knower", "Kerrie A. Herridge", "Tamara L. Howard", "Kyren A. Lazarus", "Colin D. Clyne"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0031593.g005", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/LRH_1_induces_cell_proliferation_in_17_estradiol_and_ICI_182_780_treated_cells_/352462", "title"=>"LRH-1 induces cell proliferation in 17β-estradiol and ICI 182,780 treated cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-16 00:41:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/682032"], "description"=>"<p>Quantitation of (a) LRH-1, (b) GREB1 and (c) ERα mRNA expression in estrogen-deprived MCF-7 cells (control) or LRH-1 over-expressing (+LRH-1) MCF-7 cells treated with vehicle (veh) or 10 nM 17β-estradiol (E2) for 16 h. Data is presented as mean+SE, n = 3 separate experiments, triplicate treatments per experiment, **P&lt;0.01, ***P&lt;0.001 compared to vehicle control.</p>", "links"=>[], "tags"=>["lrh-1", "greb1"], "article_id"=>352520, "categories"=>["Cancer", "Biochemistry", "Molecular Biology"], "users"=>["Ashwini L. Chand", "Dhilushi D. Wijayakumara", "Kevin C. Knower", "Kerrie A. Herridge", "Tamara L. Howard", "Kyren A. Lazarus", "Colin D. Clyne"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0031593.g006", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Synergistic_effects_of_LRH_1_and_17_estradiol_treatment_on_GREB1_expression_/352520", "title"=>"Synergistic effects of LRH-1 and 17β-estradiol treatment on GREB1 expression.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-16 00:42:00"}
  • {"files"=>["https://ndownloader.figshare.com/files/681889"], "description"=>"<p>Transcriptional activation of (a) 2×ERE and (b) GREB-ERE2 luciferase reporters by ERα and LRH-1 with vehicle (veh) or 10 nM 17β-estradiol (E2). Estrogen-deprived MCF-7 cells were over expressed with LRH-1 or ERα alone, or in combination with the appropriate reporter construct. Cells were treated with 17β-estradiol for 16 h prior to luciferase assays. Data is presented as mean+SE, n = 3 separate experiments, treatments in triplicate per experiment. *P&lt;0.05, *P&lt;0.01, ***P&lt;0.001 compared to vehicle control unless indicated by reference line.</p>", "links"=>[], "tags"=>["acts", "synergistically", "activate", "ere", "containing"], "article_id"=>352371, "categories"=>["Cancer", "Biochemistry", "Molecular Biology"], "users"=>["Ashwini L. Chand", "Dhilushi D. Wijayakumara", "Kevin C. Knower", "Kerrie A. Herridge", "Tamara L. Howard", "Kyren A. Lazarus", "Colin D. Clyne"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0031593.g004", "stats"=>{"downloads"=>1, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/LRH_1_acts_synergistically_with_ER_to_activate_ERE_containing_promoters_/352371", "title"=>"LRH-1 acts synergistically with ERα to activate ERE containing promoters.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-16 00:39:31"}
  • {"files"=>["https://ndownloader.figshare.com/files/681604"], "description"=>"<p>(a) Changes in LRH-1 mRNA and (b) protein levels in MCF-7 cells transfected with siRNA for LRH-1 (−LRH-1) or control; with pcDNA only or LRH-1-pcDNA (+LRH-1) constructs 24 h post transfection. (c) The expression levels of GREB1 in response to LRH-1 knockdown (siRNA) and over-expression (+LRH-1). Data were presented as % fold change compared to controls of the normalized expression levels, as mean ± SD, n = 3 separate experiments.</p>", "links"=>[], "tags"=>["lrh-1", "transcriptionally", "regulates"], "article_id"=>352084, "categories"=>["Cancer", "Biochemistry", "Molecular Biology"], "users"=>["Ashwini L. Chand", "Dhilushi D. Wijayakumara", "Kevin C. Knower", "Kerrie A. Herridge", "Tamara L. Howard", "Kyren A. Lazarus", "Colin D. Clyne"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0031593.g001", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Modulation_of_LRH_1_expression_in_transcriptionally_regulates_GREB1_/352084", "title"=>"Modulation of LRH-1 expression in transcriptionally regulates GREB1.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-02-16 00:34:44"}

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Relative Metric

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