Notch Activation Is Dispensable for D, L-Sulforaphane-Mediated Inhibition of Human Prostate Cancer Cell Migration
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{"title"=>"Notch Activation Is Dispensable for D, L-Sulforaphane-Mediated Inhibition of Human Prostate Cancer Cell Migration", "type"=>"journal", "authors"=>[{"first_name"=>"Eun Ryeong", "last_name"=>"Hahm", "scopus_author_id"=>"6603611510"}, {"first_name"=>"Kumar", "last_name"=>"Chandra-Kuntal", "scopus_author_id"=>"6503976609"}, {"first_name"=>"Dhimant", "last_name"=>"Desai", "scopus_author_id"=>"7102713289"}, {"first_name"=>"Shantu", "last_name"=>"Amin", "scopus_author_id"=>"7201844947"}, {"first_name"=>"Shivendra V.", "last_name"=>"Singh", "scopus_author_id"=>"35323690600"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "sgr"=>"84866116269", "doi"=>"10.1371/journal.pone.0044957", "scopus"=>"2-s2.0-84866116269", "pui"=>"365625437", "isbn"=>"1932-6203", "pmid"=>"22970326"}, "id"=>"e315b6e7-bdf5-3572-95fc-95f347fa840e", "abstract"=>"D, L-Sulforaphane (SFN), a synthetic racemic analog of broccoli constituent L-sulforaphane, is a highly promising cancer chemopreventive agent with in vivo efficacy against chemically-induced as well as oncogene-driven cancer in preclinical rodent models. Cancer chemopreventive effect of SFN is characterized by G(2)/M phase cell cycle arrest, apoptosis induction, and inhibition of cell migration and invasion. Moreover, SFN inhibits multiple oncogenic signaling pathways often hyperactive in human cancers, including nuclear factor-κB, Akt, signal transducer and activator of transcription 3, and androgen receptor. The present study was designed to determine the role of Notch signaling, which is constitutively active in many human cancers, in anticancer effects of SFN using prostate cancer cells as a model. Exposure of human prostate cancer cells (PC-3, LNCaP, and/or LNCaP-C4-2B) to SFN as well as its naturally-occurring thio-, sulfinyl-, and sulfonyl-analogs resulted in cleavage (activation) of Notch1, Notch2, and Notch4, which was accompanied by a decrease in levels of full-length Notch forms especially at the 16- and 24-hour time points. The SFN-mediated cleavage of Notch isoforms was associated with its transcriptional activation as evidenced by RBP-Jk-, HES-1A/B- and HEY-1 luciferase reporter assays. Migration of PC-3 and LNCaP cells was decreased significantly by RNA interference of Notch1 and Notch2, but not Notch4. Furthermore, SFN-mediated inhibition of PC-3 and LNCaP cell migration was only marginally affected by knockdown of Notch1 and Notch2. Strikingly, SFN administration to Transgenic Adenocarcinoma of Mouse Prostate transgenic mice failed to increase levels of cleaved Notch1, cleaved Notch2, and HES-1 proteins in vivo in prostatic intraepithelial neoplasia, well-differentiated carcinoma or poorly-differentiated prostate cancer lesions. These results indicate that Notch activation is largely dispensable for SFN-mediated inhibition of cell migration, which should be viewed as a therapeutic advantage as Notch activation is frequent in human prostate cancers.", "link"=>"http://www.mendeley.com/research/notch-activation-dispensable-d-lsulforaphanemediated-inhibition-human-prostate-cancer-cell-migration", "reader_count"=>13, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Student > Doctoral Student"=>1, "Researcher"=>4, "Student > Ph. D. Student"=>4, "Other"=>1, "Student > Master"=>2}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Student > Doctoral Student"=>1, "Researcher"=>4, "Student > Ph. D. Student"=>4, "Other"=>1, "Student > Master"=>2}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>2, "Medicine and Dentistry"=>3, "Agricultural and Biological Sciences"=>7, "Arts and Humanities"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>3}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>7}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Arts and Humanities"=>{"Arts and Humanities"=>1}}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/580333"], "description"=>"<p>(A) Representative immunohistochemical images depicting expression of cleaved Notch1, cleaved Notch2, and HES-1 proteins in the prostatic intraepithelial neoplasia (PIN), well-differentiated prostate cancer (WD), and poorly-differentiated prostate cancer (PD) in the dorsolateral prostates from TRAMP mice of the indicated groups (×200 objective magnification). (B) Quantitation of expression (combined analysis in PIN, WD, and PD) is shown as H-score (mean ± SD; n = 5). Statistical significance was determined by Student's <i>t</i>-test.</p>", "links"=>[], "tags"=>["l-sulforaphane", "cleaved", "hes-1", "proteins"], "article_id"=>250832, "categories"=>["Cancer", "Genetics", "Developmental Biology"], "users"=>["Eun-Ryeong Hahm", "Kumar Chandra-Kuntal", "Dhimant Desai", "Shantu Amin", "Shivendra V. Singh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0044957.g007", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_D_L_sulforaphane_SFN_administration_on_cleaved_Notch1_cleaved_Notch2_and_HES_1_proteins_in_vivo_/250832", "title"=>"Effect of D, L-sulforaphane (SFN) administration on cleaved Notch1, cleaved Notch2, and HES-1 proteins <i>in vivo</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 23:40:31"}
  • {"files"=>["https://ndownloader.figshare.com/files/579862"], "description"=>"<p>(A) Chemical names, common names, and chemical formulae of analogs used in the present study. Immunoblotting for cleaved Notch1, Notch2, and Notch4 using lysates from (B) PC-3, and (C) LNCaP cells after 8-hour treatment with DMSO or different analogs (10 or 20 µM). Blots were stripped and re-probed with anti-actin antibody as a loading control. Immunoblotting for each protein was done at least twice using independently prepared lysates. Numbers above band represent changes in protein levels relative to DMSO-treated control.</p>", "links"=>[], "tags"=>["analogs", "l-sulforaphane", "cleavage", "notch"], "article_id"=>250357, "categories"=>["Cancer", "Genetics", "Developmental Biology"], "users"=>["Eun-Ryeong Hahm", "Kumar Chandra-Kuntal", "Dhimant Desai", "Shantu Amin", "Shivendra V. Singh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0044957.g002", "stats"=>{"downloads"=>1, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_analogs_of_D_L_sulforaphane_on_cleavage_of_Notch_isoforms_/250357", "title"=>"Effects of analogs of D, L-sulforaphane on cleavage of Notch isoforms.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 23:37:52"}
  • {"files"=>["https://ndownloader.figshare.com/files/580257"], "description"=>"<p>(<b>SFN</b>)<b>-mediated inhibition of PC-3 cell migration.</b> (A) Immunoblotting for full-length Notch4 protein using lysates from PC-3 cells transiently transfected with a control (nonspecific) siRNA or a Notch4-targeted siRNA. (B) Representative images (Boyden chamber assay) depicting migration of PC-3 cells transfected with a control (nonspecific) siRNA or a Notch4-targeted siRNA and treated for 24 hours with DMSO or 10 μM SFN (×100 objective magnification). (C) Quantitation of PC-3 cell migration from data shown in panel B. Two to three fields on each filter were scored for cell migration under an inverted microscope. Data represent percent cell migration normalized to control siRNA-transfected cells treated with DMSO (mean ± SD, n = 6; combined data from two independent experiments each performed in triplicate). <sup>a</sup>Significantly different (<i>P</i><0.05) compared with respective DMSO-treated controls (cells transfected with control siRNA or Notch4-targeted siRNA) by one-way ANOVA followed by Bonferroni's multiple comparison test. Each experiment was performed twice.</p>", "links"=>[], "tags"=>["dispensable"], "article_id"=>250755, "categories"=>["Cancer", "Genetics", "Developmental Biology"], "users"=>["Eun-Ryeong Hahm", "Kumar Chandra-Kuntal", "Dhimant Desai", "Shantu Amin", "Shivendra V. Singh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0044957.g006", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Notch4_is_dispensable_for_D_L_sulforaphane_/250755", "title"=>"Notch4 is dispensable for D, L-sulforaphane", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 23:40:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/580182"], "description"=>"<p>(A) Immunoblotting for full-length Notch2 protein using lysates from PC-3 and LNCaP cells transiently transfected with a control (nonspecific) siRNA or a Notch2-targeted siRNA. (B) Representative images (Boyden chamber assay) depicting migration of PC-3 and LNCaP cells transfected with a control (nonspecific) siRNA or a Notch2-targeted siRNA and treated for 24 hours with DMSO or 10 μM SFN (×100 magnification). (C) Quantitation of PC-3 and LNCaP cell migration from data shown in panel B. Two to three fields on each filter were scored for cell migration under an inverted microscope. Data represent percent cell migration normalized to control siRNA transfected cells treated with DMSO (mean ± SD, n = 6; combined data from two independent experiments each performed in triplicate). (D) Analysis of histone-associated DNA fragment release into the cytosol in PC-3 and LNCaP cells transfected with a control (nonspecific) siRNA or a Notch2-targeted siRNA and treated for 24 hours with DMSO or SFN. Data represent enrichment of histone-associated DNA fragment release into the cytosol relative to control siRNA transfected cells treated with DMSO (mean ± SD, n = 6; combined data from two independent experiments each performed in triplicate). In panels C and D, significantly different (<i>P</i><0.05) <sup>a</sup>compared with respective DMSO-treated controls (cells transfected with control siRNA or Notch2-targeted siRNA), and <sup>b</sup>between control siRNA transfected cells and Notch2-targeted siRNA transfected cells by one-way ANOVA followed by Bonferroni's multiple comparison test. Each experiment was performed twice.</p>", "links"=>[], "tags"=>["notch2", "knockdown", "cellular", "responses", "l-sulforaphane"], "article_id"=>250672, "categories"=>["Cancer", "Genetics", "Developmental Biology"], "users"=>["Eun-Ryeong Hahm", "Kumar Chandra-Kuntal", "Dhimant Desai", "Shantu Amin", "Shivendra V. Singh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0044957.g005", "stats"=>{"downloads"=>0, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_Notch2_knockdown_on_cellular_responses_to_D_L_sulforaphane_SFN_/250672", "title"=>"Effect of Notch2 knockdown on cellular responses to D, L-sulforaphane (SFN).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 23:39:40"}
  • {"files"=>["https://ndownloader.figshare.com/files/580097"], "description"=>"<p>(A) Immunoblotting for full-length Notch1 protein using lysates from PC-3 and LNCaP cells transiently transfected with a control (nonspecific) siRNA or a Notch1-targeted siRNA. (B) Representative images (Boyden chamber assay) depicting migration of PC-3 and LNCaP cells transfected with a control (nonspecific) siRNA or a Notch1-targeted siRNA and treated for 24 hours with DMSO or 10 μM SFN (×100 magnification). (C) Quantitation of PC-3 and LNCaP cell migration from data shown in panel B. Two to three fields on each filter were scored for cell migration under an inverted microscope. Data represent percent cell migration normalized to control siRNA-transfected cells treated with DMSO (mean ± SD, n = 6; combined data from two independent experiments each performed in triplicate). Significantly different (<i>P</i><0.05) <sup>a</sup>compared with respective DMSO-treated controls (cells transfected with control siRNA or Notch1-targeted siRNA), and <sup>b</sup>between control siRNA transfected cells and Notch1-targeted siRNA transfected cells by one-way ANOVA followed by Bonferroni's multiple comparison test. Each experiment was performed twice.</p>", "links"=>[], "tags"=>["notch1", "l-sulforaphane", "inhibition", "pc-3", "lncap"], "article_id"=>250584, "categories"=>["Cancer", "Genetics", "Developmental Biology"], "users"=>["Eun-Ryeong Hahm", "Kumar Chandra-Kuntal", "Dhimant Desai", "Shantu Amin", "Shivendra V. Singh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0044957.g004", "stats"=>{"downloads"=>4, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_Notch1_silencing_on_D_L_sulforaphane_SFN_mediated_inhibition_of_PC_3_and_LNCaP_cell_migration_/250584", "title"=>"Effect of Notch1 silencing on D, L-sulforaphane (SFN)-mediated inhibition of PC-3 and LNCaP cell migration.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 23:39:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/580008"], "description"=>"<p>(A) Effect of SFN treatment on RBP-Jk luciferase reporter activity (a measure of transcriptional activity of Notch) in PC-3 and LNCaP cells after 8- or 24-hour treatment with DMSO or 20 µM SFN. (B) Immunofluorescence microscopic images depicting nuclear levels of HES-1 protein in PC-3 and LNCaP cells after 24-hour treatment with DMSO or 10 µM SFN (×100 objective magnification). (C) HES-1A/B luciferase reporter activity in PC-3 and LNCaP cells after 8- or 24-hour treatment with DMSO or 20 µM SFN. (D) HEY-1 (PC-3 cells) or HES-1A/B (LNCaP-C4-2B cells) luciferase reporter activity after 8- or 24-hour treatment with DMSO or 20 µM SFN. In panels A, C, and D, results shown are mean ± SD (n = 6; combined data from two independent experiments each performed in triplicate). *Significantly different (<i>P</i><0.05) compared with corresponding DMSO-treated control by Student's <i>t</i>-test (panels A, C, and D). Each experiment was performed twice.</p>", "links"=>[], "tags"=>["l-sulforaphane", "increases", "transcriptional", "notch", "prostate", "cancer"], "article_id"=>250501, "categories"=>["Cancer", "Genetics", "Developmental Biology"], "users"=>["Eun-Ryeong Hahm", "Kumar Chandra-Kuntal", "Dhimant Desai", "Shantu Amin", "Shivendra V. Singh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0044957.g003", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_D_L_Sulforaphane_SFN_increases_transcriptional_activity_of_Notch_in_prostate_cancer_cells_/250501", "title"=>"D, L-Sulforaphane (SFN) increases transcriptional activity of Notch in prostate cancer cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 23:38:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/579728"], "description"=>"<p>Immunoblotting for cleaved and full-length Notch1, Notch2, and Notch4 using lysates from (A) PC-3, (B) LNCaP, and (C) LNCaP-C4-2B cells after 8-, 16-, or 24-hour treatment with DMSO or SFN (10 or 20 µM). Blots were stripped and re-probed with anti-actin antibody as a loading control. Immunoblotting for each protein was done at least twice using independently prepared lysates. Numbers above band represent changes in protein levels relative to corresponding DMSO-treated control.</p>", "links"=>[], "tags"=>["l-sulforaphane", "causes", "cleavage", "notch4", "proteins", "pc-3", "lncap"], "article_id"=>250212, "categories"=>["Cancer", "Genetics", "Developmental Biology"], "users"=>["Eun-Ryeong Hahm", "Kumar Chandra-Kuntal", "Dhimant Desai", "Shantu Amin", "Shivendra V. Singh"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0044957.g001", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_D_L_Sulforaphane_SFN_causes_cleavage_of_Notch1_Notch2_and_Notch4_proteins_in_PC_3_and_LNCaP_cells_/250212", "title"=>"D, L-Sulforaphane (SFN) causes cleavage of Notch1, Notch2, and Notch4 proteins in PC-3 and LNCaP cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 23:37:02"}

PMC Usage Stats | Further Information

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Relative Metric

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