SMURF1 Amplification Promotes Invasiveness in Pancreatic Cancer
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{"title"=>"SMURF1 amplification promotes invasiveness in pancreatic cancer", "type"=>"journal", "authors"=>[{"first_name"=>"Kevin A.", "last_name"=>"Kwei", "scopus_author_id"=>"6507373968"}, {"first_name"=>"A. Hunter", "last_name"=>"Shain", "scopus_author_id"=>"36999600600"}, {"first_name"=>"Ryan", "last_name"=>"Bair", "scopus_author_id"=>"56103689500"}, {"first_name"=>"Kelli", "last_name"=>"Montgomery", "scopus_author_id"=>"7102499348"}, {"first_name"=>"Collins A.", "last_name"=>"Karikari", "scopus_author_id"=>"8587349800"}, {"first_name"=>"Matt", "last_name"=>"van de Rijn", "scopus_author_id"=>"7005571510"}, {"first_name"=>"Manuel", "last_name"=>"Hidalgo", "scopus_author_id"=>"7101654996"}, {"first_name"=>"Anirban", "last_name"=>"Maitra", "scopus_author_id"=>"35390791500"}, {"first_name"=>"Murali D.", "last_name"=>"Bashyam", "scopus_author_id"=>"6602984496"}, {"first_name"=>"Jonathan R.", "last_name"=>"Pollack", "scopus_author_id"=>"7101673347"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"21887346", "sgr"=>"80051943541", "doi"=>"10.1371/journal.pone.0023924", "scopus"=>"2-s2.0-80051943541", "pui"=>"362390103", "isbn"=>"1932-6203 (Electronic)\\n1932-6203 (Linking)", "issn"=>"19326203"}, "id"=>"847bebc2-7684-3a74-a52a-92ab5c241799", "abstract"=>"Pancreatic cancer is a deadly disease, and new therapeutic targets are urgently needed. We previously identified DNA amplification at 7q21-q22 in pancreatic cancer cell lines. Now, by high-resolution genomic profiling of human pancreatic cancer cell lines and human tumors (engrafted in immunodeficient mice to enrich the cancer epithelial fraction), we define a 325 Kb minimal amplicon spanning SMURF1, an E3 ubiquitin ligase and known negative regulator of transforming growth factor β (TGFβ) growth inhibitory signaling. SMURF1 amplification was confirmed in primary human pancreatic cancers by fluorescence in situ hybridization (FISH), where 4 of 95 cases (4.2%) exhibited amplification. By RNA interference (RNAi), knockdown of SMURF1 in a human pancreatic cancer line with focal amplification (AsPC-1) did not alter cell growth, but led to reduced cell invasion and anchorage-independent growth. Interestingly, this effect was not mediated through altered TGFβ signaling, assayed by transcriptional reporter. Finally, overexpression of SMURF1 (but not a catalytic mutant) led to loss of contact inhibition in NIH-3T3 mouse embryo fibroblast cells. Together, these findings identify SMURF1 as an amplified oncogene driving multiple tumorigenic phenotypes in pancreatic cancer, and provide a new druggable target for molecularly directed therapy.", "link"=>"http://www.mendeley.com/research/smurf1-amplification-promotes-invasiveness-pancreatic-cancer", "reader_count"=>21, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Researcher"=>6, "Student > Ph. D. Student"=>8, "Student > Postgraduate"=>1, "Other"=>1, "Student > Master"=>2, "Lecturer > Senior Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Researcher"=>6, "Student > Ph. D. Student"=>8, "Student > Postgraduate"=>1, "Other"=>1, "Student > Master"=>2, "Lecturer > Senior Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>9, "Agricultural and Biological Sciences"=>6, "Medicine and Dentistry"=>5, "Chemistry"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>5}, "Chemistry"=>{"Chemistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>6}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>9}}, "reader_count_by_country"=>{"United States"=>1, "India"=>1}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/374981", "https://ndownloader.figshare.com/files/375006"], "description"=>"<div><p>Pancreatic cancer is a deadly disease, and new therapeutic targets are urgently needed. We previously identified DNA amplification at 7q21-q22 in pancreatic cancer cell lines. Now, by high-resolution genomic profiling of human pancreatic cancer cell lines and human tumors (engrafted in immunodeficient mice to enrich the cancer epithelial fraction), we define a 325 Kb minimal amplicon spanning <em>SMURF1</em>, an E3 ubiquitin ligase and known negative regulator of transforming growth factor β (TGFβ) growth inhibitory signaling. <em>SMURF1</em> amplification was confirmed in primary human pancreatic cancers by fluorescence <em>in situ</em> hybridization (FISH), where 4 of 95 cases (4.2%) exhibited amplification. By RNA interference (RNAi), knockdown of SMURF1 in a human pancreatic cancer line with focal amplification (AsPC-1) did not alter cell growth, but led to reduced cell invasion and anchorage-independent growth. Interestingly, this effect was not mediated through altered TGFβ signaling, assayed by transcriptional reporter. Finally, overexpression of SMURF1 (but not a catalytic mutant) led to loss of contact inhibition in NIH-3T3 mouse embryo fibroblast cells. Together, these findings identify <em>SMURF1</em> as an amplified oncogene driving multiple tumorigenic phenotypes in pancreatic cancer, and provide a new druggable target for molecularly directed therapy.</p> </div>", "links"=>[], "tags"=>["amplification", "promotes", "invasiveness", "pancreatic", "cancer"], "article_id"=>134048, "categories"=>["Molecular Biology", "Biochemistry", "Chemistry", "Cancer", "Genetics"], "users"=>["Kevin A. Kwei", "A. Hunter Shain", "Ryan Bair", "Kelli Montgomery", "Collins A. Karikari", "Matt van de Rijn", "Manuel Hidalgo", "Anirban Maitra", "Murali D. Bashyam", "Jonathan R. Pollack"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0023924.s001", "https://dx.doi.org/10.1371/journal.pone.0023924.s002"], "stats"=>{"downloads"=>10, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_SMURF1_Amplification_Promotes_Invasiveness_in_Pancreatic_Cancer/134048", "title"=>"<em>SMURF1</em> Amplification Promotes Invasiveness in Pancreatic Cancer", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2011-08-22 01:07:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/743362"], "description"=>"<p>(<b>A</b>) A stably transduced shRNA targeting SMURF1 leads to reduced SMURF1 levels (by Western blot) compared to a non-targeting control shRNA. Residual SMURF1 levels, here normalized to GAPDH, are indicated. (<b>B</b>) Stable SMURF1 knockdown reduces anchorage-independent growth (i.e. soft agar colonies). Assay done in triplicate (mean +/− 1SD shown); *, <i>P</i><0.05 (Student's t-test).</p>", "links"=>[], "tags"=>["amplified", "aspc-1", "cells", "anchorage-independent"], "article_id"=>413722, "categories"=>["Molecular Biology", "Biochemistry", "Chemistry", "Cancer", "Genetics"], "users"=>["Kevin A. Kwei", "A. Hunter Shain", "Ryan Bair", "Kelli Montgomery", "Collins A. Karikari", "Matt van de Rijn", "Manuel Hidalgo", "Anirban Maitra", "Murali D. Bashyam", "Jonathan R. Pollack"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0023924.g003", "stats"=>{"downloads"=>5, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Knockdown_of_amplified_SMURF1_in_AsPC_1_cells_reduces_anchorage_independent_growth_/413722", "title"=>"Knockdown of amplified <i>SMURF1</i> in AsPC-1 cells reduces anchorage-independent growth.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-08-22 01:02:02"}
  • {"files"=>["https://ndownloader.figshare.com/files/743147"], "description"=>"<p>(<b>A</b>) A minimal amplicon is defined by four pancreatic cancer specimens (AsPC-1 and three xenografts). Starting from bottom: chr 7 ideogram; Heatmap of DNA copy number (red indicates gain) for the four specimens across the 7q21-q22 region (91–101 Mb); Scatter plot of DNA copy number log<sub>2</sub> ratios across 7q21.3-q22.1 (96–100 Mb), overlaid with the cghFLasso <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0023924#pone.0023924-Tibshirani1\" target=\"_blank\">[34]</a> called ratios (red line); Screen shot of the corresponding locus from the UCSC genome browser. The dashed lines bracket the 325 Kb minimal amplicon, which spans <i>SMURF1</i>. (<b>B</b>) <i>SMURF1</i> is overexpressed when gained/amplified. Box plots show 25<sup>th</sup>, 50<sup>th</sup> (median) and 75<sup>th</sup> percentile transcript levels (assayed by Agilent 44K array) for specimens with (red) or without (gray) 7q21-q22 gain, for <i>SMURF1</i> and its nearest gene neighbors. Note, <i>KPNA7</i> was not represented on the array. *, <i>P</i><0.05; **, <i>P</i><0.01; ***, <i>P</i><0.001 (Mann-Whitney U-test). (<b>C</b>) FISH reveals <i>SMURF1</i> amplification in primary pancreatic cancers. Shown are two pancreatic cancers with <i>SMURF1</i> amplification (<i>center</i>, and <i>right</i>), along with a non-amplified control (BxPC3 cells, <i>left</i>). <i>SMURF1</i> locus probe (red); control chr 7 centromere (CRP7) probe (green).</p>", "links"=>[], "tags"=>["amplification", "pancreatic", "cancer", "spans"], "article_id"=>413510, "categories"=>["Molecular Biology", "Biochemistry", "Chemistry", "Cancer", "Genetics"], "users"=>["Kevin A. Kwei", "A. Hunter Shain", "Ryan Bair", "Kelli Montgomery", "Collins A. Karikari", "Matt van de Rijn", "Manuel Hidalgo", "Anirban Maitra", "Murali D. Bashyam", "Jonathan R. Pollack"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0023924.g001", "stats"=>{"downloads"=>2, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Focal_amplification_of_7q22_1_in_pancreatic_cancer_spans_SMURF1_/413510", "title"=>"Focal amplification of 7q22.1 in pancreatic cancer spans <i>SMURF1</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-08-22 00:58:30"}
  • {"files"=>["https://ndownloader.figshare.com/files/743537"], "description"=>"<p>(<b>A</b>) Transfection of SMURF1 cDNA, or a catalytic mutant of SMURF1 (C699A) (SMURF1<sup>m</sup>), leads to overexpression (by Western blot) compared to empty vector control. SMURF1 overexpression levels, normalized to GAPDH, are indicated. (<b>B</b>) SMURF1 (but not SMURF1<sup>m</sup>) overexpression in NIH-3T3 cells leads to loss of contact inhibition (i.e. increased foci formation). Assays done in triplicate (mean +/− 1SD shown); *, <i>P</i><0.05 (Student's t-test).</p>", "links"=>[], "tags"=>["overexpression", "nih-3t3", "cells", "leads"], "article_id"=>413905, "categories"=>["Molecular Biology", "Biochemistry", "Chemistry", "Cancer", "Genetics"], "users"=>["Kevin A. Kwei", "A. Hunter Shain", "Ryan Bair", "Kelli Montgomery", "Collins A. Karikari", "Matt van de Rijn", "Manuel Hidalgo", "Anirban Maitra", "Murali D. Bashyam", "Jonathan R. Pollack"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0023924.g005", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_SMURF1_overexpression_in_NIH_3T3_cells_leads_to_loss_of_contact_inhibition_/413905", "title"=>"SMURF1 overexpression in NIH-3T3 cells leads to loss of contact inhibition.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-08-22 01:05:05"}
  • {"files"=>["https://ndownloader.figshare.com/files/743269"], "description"=>"<p>(<b>A</b>) Four different siRNAs targeting <i>SMURF1</i>, and a pool of all four, lead to reduced SMURF1 levels (by Western blot) compared to a non-targeting control siRNA pool). Residual SMURF1 levels, here normalized to GAPDH, are indicated. (<b>B</b>) SMURF1 knockdown (using siRNA pool) does not reduce cell proliferation/viability, measured by WST1 assay, and done in triplicate (mean +/− 1SD shown). (<b>C</b>) SMURF1 knockdown using four different siRNAs does not significantly alter cell proliferation/viability, measured three days post transfection. Done in triplicate (mean +/− 1SD shown). (<b>D</b>) SMURF1 knockdown does not reduce cell-cycle progression (S-phase), measured by BrdU incorporation (1.5 hr and 4 hr pulse labeling), done in triplicate (mean +/− 1 SD shown). (<b>E</b>) SMURF1 knockdown (using siRNA pool and individual siRNAs) inhibits cell invasion through Matrigel. Boyden chamber assay done in triplicate and harvested three days post transfection (mean +/− 1SD shown); *, <i>P</i><0.05 (Student's t-test). (<b>F</b>) SMURF1 knockdown does not enhance TGFβ pathway-mediated transcription. AsPC-1 cells were co-transfected with siRNAs and p3TP-Lux reporter, done in triplicate, and firefly/Renilla luciferase ratios shown (mean +/− 1SD shown). Panc1 cells (with wildtype <i>SMAD4</i>) +/− TGFβ serve as a positive control.</p>", "links"=>[], "tags"=>["knockdown", "amplified", "aspc-1", "cells"], "article_id"=>413634, "categories"=>["Molecular Biology", "Biochemistry", "Chemistry", "Cancer", "Genetics"], "users"=>["Kevin A. Kwei", "A. Hunter Shain", "Ryan Bair", "Kelli Montgomery", "Collins A. Karikari", "Matt van de Rijn", "Manuel Hidalgo", "Anirban Maitra", "Murali D. Bashyam", "Jonathan R. Pollack"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0023924.g002", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_SMURF1_knockdown_in_amplified_AsPC_1_cells_reduces_invasion_but_not_growth_/413634", "title"=>"SMURF1 knockdown in amplified AsPC-1 cells reduces invasion but not growth.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-08-22 01:00:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/743443"], "description"=>"<p>(<b>A</b>) Western blot analysis of endogenous SMURF1 protein levels in a panel of pancreatic cancer cell lines. Note that SMURF1 is highly expressed only in the 7q22.1-amplified AsPC-1 cell line. Two different exposures of the SMURF1 blot are shown; GAPDH serves as a loading control. (<b>B</b>) Western blot verification of SMURF1 knockdown (by SMURF1-targeting siRNA pool, compared to non-targeting control siRNA pool) in BxPC-3 and Hs700T cells. (<b>C</b>) Cell proliferation/viability assayed (by WST-1) in BxPC-3 cells following SMURF1 siRNA-mediated knockdown (compared to non-targeting control). Assay done in triplicate (mean +/− 1SD shown); *, <i>P</i><0.05 (Student's t-test). (<b>D</b>) Cell proliferation/viability assayed in Hs700T cells, as above. (<b>E</b>) Cell invasion assayed (by Boyden chamber) in BxPC-3 cells following SMURF1 knockdown (compared to non-targeting control). Assay done in triplicate (mean +/− 1SD shown). Note, the reduced proliferation observed with SMURF1 knockdown in Hs700T cells precluded an assay of cell invasion (where invaded cell numbers at 72 hrs are influenced by doubling times).</p>", "links"=>[], "tags"=>["smurf1", "knockdown", "pancreatic", "cancer"], "article_id"=>413809, "categories"=>["Molecular Biology", "Biochemistry", "Chemistry", "Cancer", "Genetics"], "users"=>["Kevin A. Kwei", "A. Hunter Shain", "Ryan Bair", "Kelli Montgomery", "Collins A. Karikari", "Matt van de Rijn", "Manuel Hidalgo", "Anirban Maitra", "Murali D. Bashyam", "Jonathan R. Pollack", "Alan Shain"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0023924.g004", "stats"=>{"downloads"=>2, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Analysis_of_SMURF1_knockdown_in_other_pancreatic_cancer_lines_/413809", "title"=>"Analysis of SMURF1 knockdown in other pancreatic cancer lines.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-08-22 01:03:29"}

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Relative Metric

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