Roles of PI3K/Akt and c-Jun Signaling Pathways in Human Papillomavirus Type 16 Oncoprotein-Induced HIF-1α, VEGF, and IL-8 Expression and In Vitro Angiogenesis in Non-Small Cell Lung Cancer Cells
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{"title"=>"Roles of PI3K/Akt and c-Jun signaling pathways in human papillomavirus type 16 oncoprotein-induced HIF-1α, VEGF, and IL-8 expression and in vitro angiogenesis in non-small cell lung cancer cells", "type"=>"journal", "authors"=>[{"first_name"=>"Erying", "last_name"=>"Zhang", "scopus_author_id"=>"55202031900"}, {"first_name"=>"Xiaowei", "last_name"=>"Feng", "scopus_author_id"=>"55884985800"}, {"first_name"=>"Fei", "last_name"=>"Liu", "scopus_author_id"=>"55497579300"}, {"first_name"=>"Peihua", "last_name"=>"Zhang", "scopus_author_id"=>"56291971400"}, {"first_name"=>"Jie", "last_name"=>"Liang", "scopus_author_id"=>"57198351035"}, {"first_name"=>"Xudong", "last_name"=>"Tang", "scopus_author_id"=>"57193358965"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"25058399", "issn"=>"19326203", "sgr"=>"84904812965", "pui"=>"373618177", "scopus"=>"2-s2.0-84904812965", "isbn"=>"2012010008", "doi"=>"10.1371/journal.pone.0103440"}, "id"=>"c9035017-77bd-3362-8c47-a13da1f6396e", "abstract"=>"BACKGROUND AND OBJECTIVES: Human papillomavirus (HPV)-16 infection may be related to non-smoking associated lung cancer. Our previous studies have found that HPV-16 oncoproteins promoted angiogenesis via enhancing hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF), and interleukin-8 (IL-8) expression in non-small cell lung cancer (NSCLC) cells. In this study, we further investigated the roles of PI3K/Akt and c-Jun signaling pathways in it.\\n\\nMETHODS: Human NSCLC cell lines, A549 and NCI-H460, were stably transfected with pEGFP-16 E6 or E7 plasmids. Western blotting was performed to analyze the expression of HIF-1α, p-Akt, p-P70S6K, p-P85S6K, p-mTOR, p-JNK, and p-c-Jun proteins. VEGF and IL-8 protein secretion and mRNA levels were determined by ELISA and Real-time PCR, respectively. The in vitro angiogenesis was observed by human umbilical vein endothelial cells (HUVECs) tube formation assay. Co-immunoprecipitation was performed to analyze the interaction between c-Jun and HIF-1α.\\n\\nRESULTS: HPV-16 E6 and E7 oncoproteins promoted the activation of Akt, P70S6K, P85S6K, mTOR, JNK, and c-Jun. LY294002, a PI3K inhibitor, inhibited HPV-16 oncoprotein-induced activation of Akt, P70S6K, and P85S6K, expression of HIF-1α, VEGF, and IL-8, and in vitro angiogenesis. c-Jun knockdown by specific siRNA abolished HPV-16 oncoprotein-induced HIF-1α, VEGF, and IL-8 expression and in vitro angiogenesis. Additionally, HPV-16 oncoproteins promoted HIF-1α protein stability via blocking proteasome degradation pathway, but c-Jun knockdown abrogated this effect. Furthermore, HPV-16 oncoproteins increased the quantity of c-Jun binding to HIF-1α.\\n\\nCONCLUSIONS: PI3K/Akt signaling pathway and c-Jun are involved in HPV-16 oncoprotein-induced HIF-1α, VEGF, and IL-8 expression and in vitro angiogenesis. Moreover, HPV-16 oncoproteins promoted HIF-1α protein stability possibly through enhancing the interaction between c-Jun and HIF-1α, thus making a contribution to angiogenesis in NSCLC cells.", "link"=>"http://www.mendeley.com/research/roles-pi3kakt-cjun-signaling-pathways-human-papillomavirus-type-16-oncoproteininduced-hif1%CE%B1-vegf-il8-1", "reader_count"=>23, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Researcher"=>2, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>10, "Other"=>1, "Student > Master"=>6, "Student > Bachelor"=>2}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Researcher"=>2, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>10, "Other"=>1, "Student > Master"=>6, "Student > Bachelor"=>2}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Engineering"=>2, "Biochemistry, Genetics and Molecular Biology"=>5, "Agricultural and Biological Sciences"=>10, "Medicine and Dentistry"=>5}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>2}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>5}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>10}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Unspecified"=>{"Unspecified"=>1}}, "group_count"=>0}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1608266"], "description"=>"<p>HPV-16 E6-transfected NSCLC cells were pretreated for 24 h with different concentrations of LY294002. (A) HIF-1α and p-Akt protein levels in transfected NSCLC cells (Left: A549, Right: NCI-H460) were analyzed by Western blotting. (B) VEGF and IL-8 protein concentration in the conditioned media derived from transfected A549 cells was determined by ELISA. (C) VEGF and IL-8 mRNA levels in transfected A549 cells were determined by real-time PCR. (D) HUVECs (5×10<sup>3</sup>cells/well) were seeded onto the surface of 96-well cell culture plates pre-coated with polymerized ECMatrix and then incubated at 37°C for 6 to 8 h in the conditioned media derived from HPV-16 E6-transfected A549 cells in the absence or presence of LY294002. Left: The tube formation was observed under a phase-contrast microscope (20×). Right: The total tube length in three random view-fields per well was by Scion image software measured and average value was calculated. All data are expressed as mean ± SD of three independent experiments. *<i>P</i><0.05, **<i>P</i><0.01.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "molecular biology", "Molecular cell biology", "oncology", "Basic cancer research", "Viral and bacterial causes of cancer", "Cancer risk factors", "ly294002", "hpv-16", "e6-induced", "il-8", "angiogenesis", "nsclc"], "article_id"=>1117016, "categories"=>["Biological Sciences"], "users"=>["Erying Zhang", "Xiaowei Feng", "Fei Liu", "Peihua Zhang", "Jie Liang", "Xudong Tang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0103440.g002", "stats"=>{"downloads"=>2, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_LY294002_on_HPV_16_E6_induced_HIF_1_VEGF_and_IL_8_expression_and_in_vitro_angiogenesis_in_NSCLC_cells_/1117016", "title"=>"Effect of LY294002 on HPV-16 E6-induced HIF-1α, VEGF, and IL-8 expression and <i>in vitro</i> angiogenesis in NSCLC cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-07-24 03:37:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/1608275"], "description"=>"<p>(A) HPV-16 E6- or E7-transfected NSCLC cells (Left: A549, Right: NCI-H460) were treated with 10 µg/mL of cycloheximide (CHX) for different time periods. HIF-1α protein levels were determined by Western blotting. (B<b>)</b> Quantitative densitometric analysis of results from A<b>.</b> (C) HPV-16 E6- or E7-transfected NSCLC cells (Left: A549, Right: NCI-H460) were treated with 20 µmol/L of MG-132 for 6 h. Western blotting was performed to determine HIF-1α protein levels. (D) VHL protein levels in transfected NSCLC cells (Left: A549, Right: NCI-H460) were analyzed by Western blotting. Data presented are representative of results from three independent experiments.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "molecular biology", "Molecular cell biology", "oncology", "Basic cancer research", "Viral and bacterial causes of cancer", "Cancer risk factors", "hpv-16", "e6", "e7-transfected", "nsclc"], "article_id"=>1117025, "categories"=>["Biological Sciences"], "users"=>["Erying Zhang", "Xiaowei Feng", "Fei Liu", "Peihua Zhang", "Jie Liang", "Xudong Tang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0103440.g004", "stats"=>{"downloads"=>4, "page_views"=>49, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Stability_of_HIF_1_945_protein_in_HPV_16_E6_or_E7_transfected_NSCLC_cells_/1117025", "title"=>"Stability of HIF-1α protein in HPV-16 E6 or E7-transfected NSCLC cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-07-24 03:37:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/1608279"], "description"=>"<p>(A) Western blot analysis of p-JNK, p-c-Jun, and c-Jun protein levels in stable-transfected NSCLC cells (Left: A549, Right: NCI-H460). (B and C) HPV-16 E6- or E7-transfected NSCLC cells (Left: A549, Right: NCI-H460) were pretreated with SP600125. HIF-1α, p-c-Jun, and c-Jun protein levels were analyzed by Western blotting (B), and VEGF and IL-8 protein concentration in the conditioned media was determined by ELISA (C). (D and E) HPV-16 E6- or E7-transfected NSCLC cells (Left: A549, Right: NCI-H460) were co-transfected with c-Jun siRNA (Si-1 or Si-2) or non-specific siRNA (NS-siRNA). HIF-1α, p-c-Jun, and c-Jun protein levels were analyzed by Western blotting (D), and VEGF and IL-8 protein concentration in the conditioned media was determined by ELISA (E). All data are expressed as mean ± SD of three independent experiments. *<i>P</i><0.05, **<i>P</i><0.01.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "molecular biology", "Molecular cell biology", "oncology", "Basic cancer research", "Viral and bacterial causes of cancer", "Cancer risk factors", "pathway", "il-8", "induced", "hpv-16", "oncoproteins", "nsclc"], "article_id"=>1117029, "categories"=>["Biological Sciences"], "users"=>["Erying Zhang", "Xiaowei Feng", "Fei Liu", "Peihua Zhang", "Jie Liang", "Xudong Tang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0103440.g005", "stats"=>{"downloads"=>0, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Role_of_JNK_c_Jun_signaling_pathway_in_HIF_1_945_VEGF_and_IL_8_expression_induced_by_HPV_16_oncoproteins_in_NSCLC_cells_/1117029", "title"=>"Role of JNK/c-Jun signaling pathway in HIF-1α, VEGF, and IL-8 expression induced by HPV-16 oncoproteins in NSCLC cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-07-24 03:37:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/1608281"], "description"=>"<p>HPV-16 E6- (A) or E7- (B) transfected A549 cells were co-transfected with c-Jun siRNA (Si-1 or Si-2). Left: The tube formation was observed under a phase-contrast microscope (20×). Right: The total tube length in three random view-fields per well was by Scion image software measured and average value was calculated. All data are expressed as mean ± SD of three independent experiments. **<i>P</i><0.01.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "molecular biology", "Molecular cell biology", "oncology", "Basic cancer research", "Viral and bacterial causes of cancer", "Cancer risk factors", "c-jun", "sirna", "angiogenesis", "stimulated", "over-expression", "hpv-16", "e6", "e7", "a549"], "article_id"=>1117031, "categories"=>["Biological Sciences"], "users"=>["Erying Zhang", "Xiaowei Feng", "Fei Liu", "Peihua Zhang", "Jie Liang", "Xudong Tang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0103440.g006", "stats"=>{"downloads"=>7, "page_views"=>62, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_c_Jun_siRNA_on_angiogenesis_in_vitro_stimulated_by_over_expression_of_HPV_16_E6_or_E7_in_A549_cells_/1117031", "title"=>"Effect of c-Jun siRNA on angiogenesis <i>in vitro</i> stimulated by over-expression of HPV-16 E6 or E7 in A549 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-07-24 03:37:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/1608282"], "description"=>"<p>(A) HPV-16 E6-(Left) or E7-(Right) transfected A549 and NCI-H460 cells were co-transfected with c-Jun siRNA (Si-1 or Si-2) or NS-siRNA, followed by treatment with 10 µg/mL CHX for different time periods. HIF-1α protein levels were determined by Western blotting. (B) Quantitative densitometric analysis of results from A. (C) HPV-16 E6- or E7-transfected A549 (Left) and NCI-H460 cells (Right) were co-transfected with c-Jun siRNA (Si-1 or Si-2) or NS-siRNA, followed by treatment with 20 µmol/L of MG-132 for 6 h. Western blotting was performed to determine HIF-1α protein levels and ubiquitination. (D) Co-immunoprecipitation results of HIF-1α and c-Jun in A549 cells. Upper: HPV-16 E6; Lower: HPV-16 E7. (E) Cell immunofluorescence results in A549 cells (40×). The green area showed the position of c-Jun expression in A549 cells. The red area showed the position of HIF-1α expression in A549 cells. The yellow area showed the position of HIF-1α and c-Jun co-expression. Data presented are representative of results from three independent experiments.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "molecular biology", "Molecular cell biology", "oncology", "Basic cancer research", "Viral and bacterial causes of cancer", "Cancer risk factors", "oncoproteins", "promoted", "enhancing", "c-jun"], "article_id"=>1117032, "categories"=>["Biological Sciences"], "users"=>["Erying Zhang", "Xiaowei Feng", "Fei Liu", "Peihua Zhang", "Jie Liang", "Xudong Tang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0103440.g007", "stats"=>{"downloads"=>3, "page_views"=>45, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_HPV_16_oncoproteins_promoted_HIF_1_945_protein_stability_through_enhancing_the_interaction_between_c_Jun_and_HIF_1_945_/1117032", "title"=>"HPV-16 oncoproteins promoted HIF-1α protein stability through enhancing the interaction between c-Jun and HIF-1α.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-07-24 03:37:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/1608270"], "description"=>"<p>HPV-16 E7-transfected NSCLC cells were pretreated for 24 h with different concentrations of LY294002. (A) HIF-1α and p-Akt protein levels in transfected NSCLC cells (Left: A549, Right: NCI-H460) were analyzed by Western blotting. (B) VEGF and IL-8 protein concentration in the conditioned media derived from transfected A549 cells was determined by ELISA. (C) VEGF and IL-8 mRNA levels in transfected A549 cells were determined by real-time PCR. (D) Results <i>in </i><i>vitro</i> angiogenesis (A549 cells). HUVECs (5×10<sup>3</sup>cells/well) were seeded onto the surface of 96-well cell culture plates pre-coated with polymerized ECMatrix and then incubated at 37°C for 6 to 8 h in the conditioned media derived from HPV-16 E7-transfected A549 cells in the absence or presence of LY294002. Left: The tube formation was observed under a phase-contrast microscope (20×). Right: The total tube length in 3 random view-fields per well was by Scion image software measured and average value was calculated. All data are expressed as mean ± SD of three independent experiments. *<i>P</i><0.05, **<i>P</i><0.01.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "molecular biology", "Molecular cell biology", "oncology", "Basic cancer research", "Viral and bacterial causes of cancer", "Cancer risk factors", "ly294002", "hpv-16", "e7-induced", "il-8", "angiogenesis", "nsclc"], "article_id"=>1117020, "categories"=>["Biological Sciences"], "users"=>["Erying Zhang", "Xiaowei Feng", "Fei Liu", "Peihua Zhang", "Jie Liang", "Xudong Tang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0103440.g003", "stats"=>{"downloads"=>2, "page_views"=>28, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_LY294002_on_HPV_16_E7_induced_HIF_1_VEGF_and_IL_8_expression_and_in_vitro_angiogenesis_in_NSCLC_cells_/1117020", "title"=>"Effect of LY294002 on HPV-16 E7-induced HIF-1α, VEGF, and IL-8 expression and <i>in vitro</i> angiogenesis in NSCLC cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-07-24 03:37:09"}
  • {"files"=>["https://ndownloader.figshare.com/files/1608259"], "description"=>"<p>(A) Western blot analysis of HIF-1α protein levels in stable-transfected A549 cells. (B) Real-time PCR analysis of HIF-1α mRNA levels in stable-transfected A549 cells. (C and D) Western blot analysis of p-Akt, p-P70S6K, p-P85S6K, and p-mTOR protein levels in transfected A549 (C) and NCI-H460 (D) cells.</p>", "links"=>[], "tags"=>["cell biology", "Signal transduction", "cell signaling", "molecular biology", "Molecular cell biology", "oncology", "Basic cancer research", "Viral and bacterial causes of cancer", "Cancer risk factors", "hpv-16", "oncoproteins", "pathway", "activation", "nsclc"], "article_id"=>1117013, "categories"=>["Biological Sciences"], "users"=>["Erying Zhang", "Xiaowei Feng", "Fei Liu", "Peihua Zhang", "Jie Liang", "Xudong Tang"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0103440.g001", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effects_of_HPV_16_oncoproteins_on_HIF_1_945_expression_and_PI3K_Akt_mTOR_signaling_pathway_activation_in_NSCLC_cells_/1117013", "title"=>"Effects of HPV-16 oncoproteins on HIF-1α expression and PI3K/Akt/mTOR signaling pathway activation in NSCLC cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-07-24 03:37:09"}

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Relative Metric

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