Proline-Rich Tyrosine Kinase 2 (Pyk2) Regulates IGF-I-Induced Cell Motility and Invasion of Urothelial Carcinoma Cells
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{"title"=>"Proline-rich tyrosine kinase 2 (Pyk2) regulates IGF-I-induced cell motility and invasion of urothelial carcinoma cells", "type"=>"journal", "authors"=>[{"first_name"=>"Marco", "last_name"=>"Genua", "scopus_author_id"=>"14119657500"}, {"first_name"=>"Shi Qiong", "last_name"=>"Xu", "scopus_author_id"=>"7404439025"}, {"first_name"=>"Simone", "last_name"=>"Buraschi", "scopus_author_id"=>"36702435900"}, {"first_name"=>"Stephen C.", "last_name"=>"Peiper", "scopus_author_id"=>"7006043424"}, {"first_name"=>"Leonard G.", "last_name"=>"Gomella", "scopus_author_id"=>"7005666942"}, {"first_name"=>"Antonino", "last_name"=>"Belfiore", "scopus_author_id"=>"35568609400"}, {"first_name"=>"Renato V.", "last_name"=>"Iozzo", "scopus_author_id"=>"35511430300"}, {"first_name"=>"Andrea", "last_name"=>"Morrione", "scopus_author_id"=>"6701669179"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"scopus"=>"2-s2.0-84862978950", "pui"=>"365122056", "issn"=>"19326203", "doi"=>"10.1371/journal.pone.0040148", "pmid"=>"22859931", "sgr"=>"84862978950", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)"}, "id"=>"a2e66b4f-3754-3fdd-af15-45886e4ba8e0", "abstract"=>"The insulin-like growth factor receptor I (IGF-IR) plays an essential role in transformation by promoting cell growth and protecting cancer cells from apoptosis. We have recently demonstrated that the IGF-IR is overexpressed in invasive bladder cancer tissues and promotes motility and invasion of urothelial carcinoma cells. These effects require IGF-I-induced Akt- and MAPK-dependent activation of paxillin. The latter co-localizes with focal adhesion kinases (FAK) at dynamic focal adhesions and is critical for promoting motility of urothelial cancer cells. FAK and its homolog Proline-rich tyrosine kinase 2 (Pyk2) modulate paxillin activation; however, their role in regulating IGF-IR-dependent signaling and motility in bladder cancer has not been established. In this study we demonstrate that FAK was not required for IGF-IR-dependent signaling and motility of invasive urothelial carcinoma cells. On the contrary, Pyk2, which was strongly activated by IGF-I, was critical for IGF-IR-dependent motility and invasion and regulated IGF-I-dependent activation of the Akt and MAPK pathways. Using immunofluorescence and AQUA analysis we further discovered that Pyk2 was overexpressed in bladder cancer tissues as compared to normal tissue controls. Significantly, in urothelial carcinoma tissues there was increased Pyk2 localization in the nuclei as compared to normal tissue controls. These results provide the first evidence of a specific Pyk2 activity in regulating IGF-IR-dependent motility and invasion of bladder cancer cells suggesting that Pyk2 and the IGF-IR may play a critical role in the invasive phenotype in urothelial neoplasia. In addition, Pyk2 and the IGF-IR may serve as novel biomarkers with diagnostic and prognostic significance in bladder cancer.", "link"=>"http://www.mendeley.com/research/prolinerich-tyrosine-kinase-2-pyk2-regulates-igfiinduced-cell-motility-invasion-urothelial-carcinoma", "reader_count"=>12, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Librarian"=>1, "Student > Ph. D. Student"=>3, "Student > Postgraduate"=>2, "Student > Master"=>2, "Student > Bachelor"=>1, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Librarian"=>1, "Student > Ph. D. Student"=>3, "Student > Postgraduate"=>2, "Student > Master"=>2, "Student > Bachelor"=>1, "Professor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>5, "Medicine and Dentistry"=>2, "Social Sciences"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Social Sciences"=>{"Social Sciences"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>5}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>2}}, "reader_count_by_country"=>{"Denmark"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/321151", "https://ndownloader.figshare.com/files/321175"], "description"=>"<div><p>The insulin-like growth factor receptor I (IGF-IR) plays an essential role in transformation by promoting cell growth and protecting cancer cells from apoptosis. We have recently demonstrated that the IGF-IR is overexpressed in invasive bladder cancer tissues and promotes motility and invasion of urothelial carcinoma cells. These effects require IGF-I-induced Akt- and MAPK-dependent activation of paxillin. The latter co-localizes with focal adhesion kinases (FAK) at dynamic focal adhesions and is critical for promoting motility of urothelial cancer cells. FAK and its homolog Proline-rich tyrosine kinase 2 (Pyk2) modulate paxillin activation; however, their role in regulating IGF-IR-dependent signaling and motility in bladder cancer has not been established. In this study we demonstrate that FAK was not required for IGF-IR-dependent signaling and motility of invasive urothelial carcinoma cells. On the contrary, Pyk2, which was strongly activated by IGF-I, was critical for IGF-IR-dependent motility and invasion and regulated IGF-I-dependent activation of the Akt and MAPK pathways. Using immunofluorescence and AQUA analysis we further discovered that Pyk2 was overexpressed in bladder cancer tissues as compared to normal tissue controls. Significantly, in urothelial carcinoma tissues there was increased Pyk2 localization in the nuclei as compared to normal tissue controls. These results provide the first evidence of a specific Pyk2 activity in regulating IGF-IR-dependent motility and invasion of bladder cancer cells suggesting that Pyk2 and the IGF-IR may play a critical role in the invasive phenotype in urothelial neoplasia. In addition, Pyk2 and the IGF-IR may serve as novel biomarkers with diagnostic and prognostic significance in bladder cancer.</p> </div>", "links"=>[], "tags"=>["proline-rich", "tyrosine", "kinase", "regulates", "igf-i-induced", "motility", "urothelial", "carcinoma", "cells"], "article_id"=>123377, "categories"=>["Cancer", "Molecular Biology", "Marine Biology", "Cell Biology"], "users"=>["Marco Genua", "Shi-Qiong Xu", "Simone Buraschi", "Stephen C. Peiper", "Leonard G. Gomella", "Antonino Belfiore", "Renato V. Iozzo", "Andrea Morrione"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0040148.s001", "https://dx.doi.org/10.1371/journal.pone.0040148.s002"], "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Proline_Rich_Tyrosine_Kinase_2_Pyk2_Regulates_IGF_I_Induced_Cell_Motility_and_Invasion_of_Urothelial_Carcinoma_Cells/123377", "title"=>"Proline-Rich Tyrosine Kinase 2 (Pyk2) Regulates IGF-I-Induced Cell Motility and Invasion of Urothelial Carcinoma Cells", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-06-28 00:56:17"}
  • {"files"=>["https://ndownloader.figshare.com/files/617807"], "description"=>"<p>(A) Pyk2 expression on a bladder cancer tissue microarray was determined by immunofluorescence and AQUA analysis using the AQUA PM-2000 system (HistoRx, Inc). Automated quantification and statistics on the different types of bladder cancer tumor tissues (B) and in the cytoplasmic and nuclear fractions of urothelial carcinoma cells (C) was calculated by AQUA Software. (B) *<i>P</i><0.05. **<i>P</i><0.01 compared to normal tissue controls. (C) *<i>P</i> = 0.012 compared to non-neoplastic nuclear fraction.</p>", "links"=>[], "tags"=>["up-regulated", "bladder", "cancer"], "article_id"=>288290, "categories"=>["Cancer", "Molecular Biology", "Marine Biology", "Cell Biology"], "users"=>["Marco Genua", "Shi-Qiong Xu", "Simone Buraschi", "Stephen C. Peiper", "Leonard G. Gomella", "Antonino Belfiore", "Renato V. Iozzo", "Andrea Morrione"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0040148.g005", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Pyk2_is_up_regulated_in_bladder_cancer_tissues_/288290", "title"=>"Pyk2 is up-regulated in bladder cancer tissues.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-28 02:18:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/617460"], "description"=>"<p>(<i>A</i>) Serum-starved 5637 cells were stimulated with 50 ng/ml of IGF-I for the indicated time points. Pyk2 phosphorylation was detected by immunoblot using anti-phospho-Pyk2 (Tyr402) antibodies, while total Pyk2 protein level was assessed using anti-Pyk2 polyclonal antibodies. Blot is representative of two independent experiments. (<i>B</i>) Migration of 5637 cells transiently transfected with either Flag-tagged wild type (PYK2 WT) or a dominant negative (KD PYK2) Pyk2 proteins was assessed after 16 hours of IGF-I stimulation. Values are expressed as fold change over SFM and represent mean ± SD. ** <i>P</i><0.01. (<i>C</i>) Expression levels of transiently transfected Pyk2 proteins were assessed by immunoblot with anti-flag M2 antibodies. Blot is representative of two independent experiments.</p>", "links"=>[], "tags"=>["pyk2", "igf-ir-dependent", "motility", "invasive", "urothelial", "cancer"], "article_id"=>287942, "categories"=>["Cancer", "Molecular Biology", "Marine Biology", "Cell Biology"], "users"=>["Marco Genua", "Shi-Qiong Xu", "Simone Buraschi", "Stephen C. Peiper", "Leonard G. Gomella", "Antonino Belfiore", "Renato V. Iozzo", "Andrea Morrione"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0040148.g002", "stats"=>{"downloads"=>0, "page_views"=>40, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_IGF_I_activated_Pyk2_is_critical_for_IGF_IR_dependent_motility_of_invasive_urothelial_cancer_cells_/287942", "title"=>"IGF-I-activated Pyk2 is critical for IGF-IR-dependent motility of invasive urothelial cancer cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-28 02:12:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/617362"], "description"=>"<p>(A) 5637 cells were transfected with the FAK siGenome pool or control oligos. After 72 hours FAK and Pyk2 expression was detected by immunoblot with specific antibodies. Blot is representative of three independent experiments with an average FAK depletion level of 93.3±3.5 (arbitrary units) as assessed by densitometric analysis (B and C) Migration and invasion assays of 5637 cells were performed as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040148#s4\" target=\"_blank\">Materials and Methods</a> and assessed after 16 hours of IGF-I stimulation. Values are expressed as fold change over SFM and represent mean ± SD. *<i>P</i> = 0.046. (<i>D</i>) FAK-depleted 5637 cells were tested for Akt and MAPK activation after 10 minutes of IGF-I stimulation using a mix of phospho-specific antibodies (PathScan Cocktail I). eIF4E monitors protein loads. Blot is representative of three independent experiments.</p>", "links"=>[], "tags"=>["igf-i-mediated", "motility", "signaling", "invasive", "urothelial", "cancer"], "article_id"=>287846, "categories"=>["Cancer", "Molecular Biology", "Marine Biology", "Cell Biology"], "users"=>["Marco Genua", "Shi-Qiong Xu", "Simone Buraschi", "Stephen C. Peiper", "Leonard G. Gomella", "Antonino Belfiore", "Renato V. Iozzo", "Andrea Morrione"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0040148.g001", "stats"=>{"downloads"=>1, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_FAK_is_not_important_for_IGF_I_mediated_motility_and_signaling_of_invasive_urothelial_cancer_cells_/287846", "title"=>"FAK is not important for IGF-I-mediated motility and signaling of invasive urothelial cancer cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-28 02:10:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/617524"], "description"=>"<p>(A) 5637 cells were transfected with the Pyk2 siGenome pool or control. After 72 hours Pyk2 and FAK expression was detected by immunoblot with specific antibodies. Blot is representative of three independent experiments with an average Pyk2 depletion level of 92.6±3 (arbitrary units) as assessed by densitometric analysis (B and C) Migration and invasion of 5637 cells were assessed as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040148#s4\" target=\"_blank\">Materials and Methods</a> after 16 hours of IGF-I stimulation. Values are expressed as fold change over SFM and represent mean ± SD. *<i>P</i><0.05; **<i>P</i><0.01. (<i>D</i>) Pyk2-depleted 5637 cells were tested for the activation of the Akt and MAPK pathways after 10 min of IGF-I stimulation using a mix of phospho-specific antibodies (PathScan Cocktail I). eIF4E monitors protein loads. Blot is representative of three independent experiments.</p>", "links"=>[], "tags"=>["igf-ir-induced", "signaling", "invasive", "urothelial", "cancer"], "article_id"=>288008, "categories"=>["Cancer", "Molecular Biology", "Marine Biology", "Cell Biology"], "users"=>["Marco Genua", "Shi-Qiong Xu", "Simone Buraschi", "Stephen C. Peiper", "Leonard G. Gomella", "Antonino Belfiore", "Renato V. Iozzo", "Andrea Morrione"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0040148.g003", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Pyk2_is_critical_for_IGF_IR_induced_motility_invasion_and_signaling_of_invasive_urothelial_cancer_cells_/288008", "title"=>"Pyk2 is critical for IGF-IR-induced motility, invasion and signaling of invasive urothelial cancer cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-28 02:13:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/617630"], "description"=>"<p>(A) 5637 cells were serum-starved over night and then treated with 50 ng/ml of IGF-I for 30 minutes. After fixation, cells were labeled with a monoclonal anti-IGF-IR (green) and a polyclonal anti-Pyk2 (red) and imaged by confocal microscopy. The pictures of merged fields show colocalization (yellow) of IGF-IR and Pyk2 in the IGF-I treated cells (arrows) but not in unstimulated control cells. The distinct co-localization of Pyk2 and IGF-IR is detectable in the Z stacks (yellow staining, bottom panel). Pictures are representative of at least 10 independent fields from two independent experiments. An average of 300 cells was examined for each condition. Bar: 10 µm. (B) 5637 and(C) T24 bladder cancer cells were serum-starved for 24 hours and then stimulated with 50 ng/ml of IGF-I for 30 minutes. Two mg of cell lysates were immunoprecipitated with anti-Pyk2 polyclonal antibodies. IRS-1, IRS-2, Grb2 and Pyk2 levels were assessed by immunoblot with specific polyclonal antibodies. Blots are representatives of three independent experiments.</p>", "links"=>[], "tags"=>["colocalizes", "igf-ir", "complexes", "irs-2", "grb2", "igf-i", "stimulation", "urothelial", "cancer"], "article_id"=>288110, "categories"=>["Cancer", "Molecular Biology", "Marine Biology", "Cell Biology"], "users"=>["Marco Genua", "Shi-Qiong Xu", "Simone Buraschi", "Stephen C. Peiper", "Leonard G. Gomella", "Antonino Belfiore", "Renato V. Iozzo", "Andrea Morrione"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0040148.g004", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Pyk2_colocalizes_with_the_IGF_IR_and_complexes_with_IRS_2_and_Grb2_after_IGF_I_stimulation_of_urothelial_cancer_cells_/288110", "title"=>"Pyk2 colocalizes with the IGF-IR and complexes with IRS-2 and Grb2 after IGF-I stimulation of urothelial cancer cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-28 02:15:10"}

PMC Usage Stats | Further Information

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Relative Metric

{"start_date"=>"2012-01-01T00:00:00Z", "end_date"=>"2012-12-31T00:00:00Z", "subject_areas"=>[{"subject_area"=>"/Biology and life sciences/Biochemistry", "average_usage"=>[316, 541, 663, 766, 856, 950, 1041, 1128, 1218, 1302, 1382, 1456, 1526, 1593, 1657, 1729, 1796, 1862, 1930, 1999, 2065, 2132, 2202, 2261, 2319]}, {"subject_area"=>"/Biology and life sciences/Cell biology", "average_usage"=>[319, 556, 679, 785, 881, 970, 1062, 1149, 1236, 1323, 1402, 1474, 1545, 1617, 1681, 1754, 1822, 1892, 1963, 2031, 2099, 2165, 2233, 2299, 2359]}, {"subject_area"=>"/Biology and life sciences/Developmental biology", "average_usage"=>[313, 562, 690, 796, 893, 986, 1079, 1173, 1257, 1349, 1429, 1506, 1586, 1661, 1730, 1803, 1876, 1945, 2017, 2091, 2163, 2225, 2294, 2362, 2430]}, {"subject_area"=>"/Medicine and health sciences", "average_usage"=>[312, 547, 668, 769, 861, 953, 1046, 1135, 1224, 1307, 1388, 1464, 1536, 1606, 1676, 1744, 1812, 1882, 1954, 2020, 2089, 2155, 2218, 2282, 2344]}, {"subject_area"=>"/Medicine and health sciences/Oncology", "average_usage"=>[301, 551, 686, 790, 883, 972, 1069, 1162, 1250, 1341, 1426, 1503, 1571, 1634, 1701, 1779, 1848, 1919, 1986, 2052, 2114, 2183, 2253, 2301, 2367]}]}
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