Regulation of HIF-1α and VEGF by miR-20b Tunes Tumor Cells to Adapt to the Alteration of Oxygen Concentration
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{"title"=>"Regulation of HIF-1α and VEGF by miR-20b tunes tumor cells to adapt to the alteration of oxygen concentration", "type"=>"journal", "authors"=>[{"first_name"=>"Zhang", "last_name"=>"Lei", "scopus_author_id"=>"35262133900"}, {"first_name"=>"Bo", "last_name"=>"Li", "scopus_author_id"=>"35285845800"}, {"first_name"=>"Zhuoshun", "last_name"=>"Yang", "scopus_author_id"=>"35263053400"}, {"first_name"=>"Haoshu", "last_name"=>"Fang", "scopus_author_id"=>"35261514500"}, {"first_name"=>"Gui Mei", "last_name"=>"Zhang", "scopus_author_id"=>"7405269772"}, {"first_name"=>"Zuo Hua", "last_name"=>"Feng", "scopus_author_id"=>"7403442876"}, {"first_name"=>"Bo", "last_name"=>"Huang", "scopus_author_id"=>"55086047400"}], "year"=>2009, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "pui"=>"355633889", "doi"=>"10.1371/journal.pone.0007629", "sgr"=>"70449602451", "scopus"=>"2-s2.0-70449602451", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"19893619"}, "id"=>"bf23b1b3-7620-3ff1-ae1d-24760b239120", "abstract"=>"The regulation of HIF-1alpha is considered to be realized by pVHL-mediated ubiquitin-26S proteasome pathway at a post-transcriptional level. The discovery of a class of small noncoding RNAs, called microRNAs, implies alternative mechanism of regulation of HIF-1alpha. Here, we show that miR-20b plays an important role in fine-tuning the adaptation of tumor cells to oxygen concentration. The inhibition of miR-20b increased the protein levels of HIF-1alpha and VEGF in normoxic tumor cells; the increase of miR-20b in hypoxic tumor cells, nevertheless, decreased the protein levels of HIF-1alpha and VEGF. By using luciferase reporter vector system, we confirmed that miR-20b directly targeted the 3'UTR of Hif1a and Vegfa. On the other hand, the forced overexpression of HIF-1alpha in normoxic tumor cells downregulated miR-20b expression. However, HIF-1alpha knockdown in hypoxic tumor cells caused the increase of miR-20b. The differential expression of miR-20b has important biological significance in tumor cells, either enhancing the growth or favoring the survival of tumor cells upon the oxygen supply. Thus, we identify a novel molecular regulation mechanism through which miR-20b regulates HIF-1alpha and VEGF and is regulated by HIF-1alpha so to keep tumor cells adapting to different oxygen concentrations.", "link"=>"http://www.mendeley.com/research/regulation-hif1%CE%B1-vegf-mir20b-tunes-tumor-cells-adapt-alteration-oxygen-concentration", "reader_count"=>52, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>3, "Researcher"=>15, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>13, "Student > Postgraduate"=>1, "Student > Master"=>10, "Student > Bachelor"=>3, "Professor"=>4}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>3, "Researcher"=>15, "Student > Doctoral Student"=>3, "Student > Ph. D. Student"=>13, "Student > Postgraduate"=>1, "Student > Master"=>10, "Student > Bachelor"=>3, "Professor"=>4}, "reader_count_by_subject_area"=>{"Engineering"=>2, "Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>34, "Medicine and Dentistry"=>8, "Chemical Engineering"=>1, "Chemistry"=>1, "Computer Science"=>2}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>2}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>8}, "Chemistry"=>{"Chemistry"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>34}, "Computer Science"=>{"Computer Science"=>2}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>1}, "Chemical Engineering"=>{"Chemical Engineering"=>1}}, "reader_count_by_country"=>{"Austria"=>1, "United States"=>1, "Japan"=>1, "Italy"=>1, "Switzerland"=>1, "Spain"=>1}, "group_count"=>1}

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  • {"files"=>["https://ndownloader.figshare.com/files/435123", "https://ndownloader.figshare.com/files/435156", "https://ndownloader.figshare.com/files/435188", "https://ndownloader.figshare.com/files/435216"], "description"=>"<div><p>The regulation of HIF-1α is considered to be realized by pVHL-mediated ubiquitin-26S proteasome pathway at a post-transcriptional level. The discovery of a class of small noncoding RNAs, called microRNAs, implies alternative mechanism of regulation of HIF-1α. Here, we show that miR-20b plays an important role in fine-tuning the adaptation of tumor cells to oxygen concentration. The inhibition of miR-20b increased the protein levels of HIF-1α and VEGF in normoxic tumor cells; the increase of miR-20b in hypoxic tumor cells, nevertheless, decreased the protein levels of HIF-1α and VEGF. By using luciferase reporter vector system, we confirmed that miR-20b directly targeted the 3′UTR of <em>Hif1a</em> and <em>Vegfa</em>. On the other hand, the forced overexpression of HIF-1α in normoxic tumor cells downregulated miR-20b expression. However, HIF-1α knockdown in hypoxic tumor cells caused the increase of miR-20b. The differential expression of miR-20b has important biological significance in tumor cells, either enhancing the growth or favoring the survival of tumor cells upon the oxygen supply. Thus, we identify a novel molecular regulation mechanism through which miR-20b regulates HIF-1α and VEGF and is regulated by HIF-1α so to keep tumor cells adapting to different oxygen concentrations.</p></div>", "links"=>[], "tags"=>["vegf", "mir-20b", "tunes", "cells", "alteration"], "article_id"=>145833, "categories"=>["Cancer", "Cell Biology", "Molecular Biology"], "users"=>["Zhang Lei", "Bo Li", "Zhuoshun Yang", "Haoshu Fang", "Gui-Mei Zhang", "Zuo-Hua Feng", "Bo Huang"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0007629.s001", "https://dx.doi.org/10.1371/journal.pone.0007629.s002", "https://dx.doi.org/10.1371/journal.pone.0007629.s003", "https://dx.doi.org/10.1371/journal.pone.0007629.s004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Regulation_of_HIF_1_and_VEGF_by_miR_20b_Tunes_Tumor_Cells_to_Adapt_to_the_Alteration_of_Oxygen_Concentration/145833", "title"=>"Regulation of HIF-1α and VEGF by miR-20b Tunes Tumor Cells to Adapt to the Alteration of Oxygen Concentration", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2009-10-29 01:37:13"}
  • {"files"=>["https://ndownloader.figshare.com/files/878073"], "description"=>"<p>(A) The expressions of miR-18a, 155, 20b and 199b in tumor cell lines were detected by RT-PCR in normoxia (20% oxygen) or hypoxia (1% oxygen). (B) The detection of miR-20b in tumor cell lines by real time RT-PCR. The miR-20b level in the hypoxia groups was designated as 1. (C and D) The expression of HIF-1α in tumor cell lines was detected by Western blot (C) or real time RT-PCR (D). (E) Expressions of miR-20b and HIF-1α in different tumor regions. 1×10<sup>5</sup> H22 tumor cells were inoculated subcutaneously into mice. When tumor size was >9×9 mm, the central and marginal regions of tumor tissues were used for miR-20b and HIF-1α detection by RT-PCR and Western blot, respectively. Data from three tumor tissues were presented in this figure.</p>", "links"=>[], "tags"=>["mir-20b", "reversely", "correlated"], "article_id"=>548523, "categories"=>["Cancer", "Cell Biology", "Molecular Biology", "Virology"], "users"=>["Zhang Lei", "Bo Li", "Zhuoshun Yang", "Haoshu Fang", "Gui-Mei Zhang", "Zuo-Hua Feng", "Bo Huang"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0007629.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_of_miR_20b_is_reversely_correlated_with_HIF_1_945_level_in_tumor_cells_/548523", "title"=>"Expression of miR-20b is reversely correlated with HIF-1α level in tumor cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2009-10-29 02:22:03"}
  • {"files"=>["https://ndownloader.figshare.com/files/878144"], "description"=>"<p>(A) miR-20b inhibitor increased HIF-1α protein in normoxic H22 cells. miR-20b inhibitor or control oligonucleotide was transfected into normoxic H22 cells. 48 hr later, the cells were used to detected HIF-1α and VHL proteins by Western blot. (B) miR-20b decreased HIF-1α protein in hypoxic H22 cells. miR-20b or control oligonucleotide was transfected into hypoxic H22 cells. 48 hr later, the cells were used to detect HIF-1α and VHL proteins by Western blot. (C) miR-20b inhibitor or miR-20b did not affect the mRNA levels of HIF-1α. The cells of above (A) and (B) were used for real time RT-PCR to detect HIF-1α mRNA expression. (D) miR-20b targeted 3′-UTR of HIF-1α mRNA. The specificity of miR-20b to 3′-UTR of HIF-1α mRNA was identified as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007629#s4\" target=\"_blank\"><i>Methods</i></a>. *, <i>P</i><0.01, compared with HIF-1α 3′-UTR group.</p>", "links"=>[], "tags"=>["targeted"], "article_id"=>548602, "categories"=>["Cancer", "Cell Biology", "Molecular Biology", "Virology"], "users"=>["Zhang Lei", "Bo Li", "Zhuoshun Yang", "Haoshu Fang", "Gui-Mei Zhang", "Zuo-Hua Feng", "Bo Huang"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0007629.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_HIF_1_945_is_targeted_by_miR_20b_/548602", "title"=>"HIF-1α is targeted by miR-20b.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2009-10-29 02:23:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/878211"], "description"=>"<p>(A and B) HIF-1α decreased miR-20b expression in normoxic H22 cells. HIF-1α vector or mock vector was transfected into normoxic H22 cells. 72 hr later, the cells were used for HIF-1α analysis by Western blot (A) and miR-20b analysis by RT-PCR and real time RT-PCR (B). (C and D) HIF-1α siRNA was transfected into hypoxic H22 cells. 72 hr later, the cells were used for HIF-1α analysis by Western blot (C) and miR-20b analysis by RT-PCR and real time RT-PCR (D). (E and F) VHL siRNA was transfected into normoxic H22 cells. 72 hr later, the cells were used for VHL and HIF-1α analysis by Western blot (E) and miR-20b analysis by RT-PCR and real time RT-PCR (F).</p>", "links"=>[], "tags"=>["downregulated"], "article_id"=>548666, "categories"=>["Cancer", "Cell Biology", "Molecular Biology", "Virology"], "users"=>["Zhang Lei", "Bo Li", "Zhuoshun Yang", "Haoshu Fang", "Gui-Mei Zhang", "Zuo-Hua Feng", "Bo Huang"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0007629.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_miR_20b_is_downregulated_by_HIF_1_945_/548666", "title"=>"miR-20b is downregulated by HIF-1α.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2009-10-29 02:24:26"}
  • {"files"=>["https://ndownloader.figshare.com/files/878297"], "description"=>"<p>(A) miR-20b inhibitor increased VEGF protein in normoxic H22 cells. miR-20b inhibitor or control oligonucleotide was transfected into normoxic H22 cells. 48 hr later, the cells were used to detected VEGF expression by Western blot. (B) miR-20b decreased VEGF protein in hypoxic H22 cells. miR-20b or control oligonucleotide was transfected into hypoxic H22 cells. 48 hr later, the cells were used to detect VEGF expression by Western blot. (C) miR-20b inhibitor or miR-20b did not affect the mRNA levels of VEGF. The cells of above (A) and (B) were used for real time RT-PCR to detect VEGF mRNA expression. (D) miR-20b targeted 3′-UTR of VEGF mRNA. The specificity of miR-20b to 3′-UTR of VEGF mRNA was identified as described in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0007629#s4\" target=\"_blank\"><i>Methods</i></a>. *, <i>P</i><0.01, compared with VEGF 3′-UTR group.</p>", "links"=>[], "tags"=>["targeted"], "article_id"=>548752, "categories"=>["Cancer", "Cell Biology", "Molecular Biology", "Virology"], "users"=>["Zhang Lei", "Bo Li", "Zhuoshun Yang", "Haoshu Fang", "Gui-Mei Zhang", "Zuo-Hua Feng", "Bo Huang"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0007629.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_VEGF_is_targeted_by_miR_20b_/548752", "title"=>"VEGF is targeted by miR-20b.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2009-10-29 02:25:52"}
  • {"files"=>["https://ndownloader.figshare.com/files/878362"], "description"=>"<p>(A) miR-20b was required for H22 cell growth. H22 cells were transfected with miR-20b inhibitor or miR-20b inhibitor + HIF-1α siRNA or miR-20b or VHL siRNA or control oligonucleotide for 24 hr. Then the cells were seeded in 96-well plate (5×10<sup>3</sup> per well) for 48 h in normoxia. The proliferation assay was performed with MTT Cell Proliferation Kit (Roche Diagnostics, IN) according to the manufacturer's instructions. *, <i>P</i><0.05, compared with control. (B) Downregulation of miR-20b enhanced the resistance to apotosis. H22 cells were transfected with miR-20b inhibitor or miR-20b inhibitor + HIF-1α siRNA for 24 hr. Then the cells (1×10<sup>6</sup>) were irradiated by UVB (200 J/m<sup>2</sup>) or treated with mitomycin C (MMC, 10 µg/ml)for 12 h. The cells were stained with PE-Annexin V and 7-AAD for apoptotic analysis by flow cytometry. *, <i>P</i><0.05, compared with control. (C) Analysis of the mRNA expressions of Bcl-2, Bcl-xL, Bax and Bad genes. H22 cells were transfected with miR-20b inhibitor or miR-20b inhibitor + HIF-1α siRNA for 72 hr. The cells were used for the analysis of gene expression by real time RT-PCR. *, <i>P</i><0.05, compared with control. (D) Normoxic H22 cells were transfected with miR-20b for 24 hr. Then the cells were cultured under hypoxic condition for 12 h, and stained with PE-Annexin V and 7-AAD for apoptotic analysis by flow cytometry. *, <i>P</i><0.05, compared with control.</p>", "links"=>[], "tags"=>["mir-20b", "affects", "aspects"], "article_id"=>548811, "categories"=>["Cancer", "Cell Biology", "Molecular Biology", "Virology"], "users"=>["Zhang Lei", "Bo Li", "Zhuoshun Yang", "Haoshu Fang", "Gui-Mei Zhang", "Zuo-Hua Feng", "Bo Huang"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0007629.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Differential_expression_of_miR_20b_affects_different_aspects_of_tumor_cells_/548811", "title"=>"Differential expression of miR-20b affects different aspects of tumor cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2009-10-29 02:26:51"}

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  • {"unique-ip"=>"23", "full-text"=>"29", "pdf"=>"6", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"27", "supp-data"=>"3", "cited-by"=>"0", "year"=>"2016", "month"=>"11"}
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  • {"unique-ip"=>"11", "full-text"=>"11", "pdf"=>"4", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"11", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"9"}
  • {"unique-ip"=>"8", "full-text"=>"9", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"12", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"10"}
  • {"unique-ip"=>"12", "full-text"=>"14", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"28", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"11"}
  • {"unique-ip"=>"8", "full-text"=>"7", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"12"}
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  • {"unique-ip"=>"14", "full-text"=>"11", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"10", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"3"}
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  • {"unique-ip"=>"5", "full-text"=>"4", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"6"}
  • {"unique-ip"=>"10", "full-text"=>"10", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"12", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"9"}
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  • {"unique-ip"=>"18", "full-text"=>"19", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"11", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"12"}
  • {"unique-ip"=>"10", "full-text"=>"12", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"12", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2018", "month"=>"10"}
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  • {"unique-ip"=>"12", "full-text"=>"12", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2019", "month"=>"8"}
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  • {"unique-ip"=>"17", "full-text"=>"17", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"9", "supp-data"=>"4", "cited-by"=>"0", "year"=>"2019", "month"=>"10"}
  • {"unique-ip"=>"14", "full-text"=>"11", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"15", "supp-data"=>"4", "cited-by"=>"0", "year"=>"2019", "month"=>"12"}
  • {"unique-ip"=>"17", "full-text"=>"18", "pdf"=>"4", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"4", "cited-by"=>"0", "year"=>"2020", "month"=>"2"}
  • {"unique-ip"=>"16", "full-text"=>"20", "pdf"=>"5", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"4", "cited-by"=>"0", "year"=>"2020", "month"=>"3"}
  • {"unique-ip"=>"6", "full-text"=>"5", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"4"}
  • {"unique-ip"=>"2", "full-text"=>"2", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"5"}
  • {"unique-ip"=>"8", "full-text"=>"9", "pdf"=>"2", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"6"}
  • {"unique-ip"=>"5", "full-text"=>"4", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"7"}
  • {"unique-ip"=>"6", "full-text"=>"5", "pdf"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"8"}
  • {"unique-ip"=>"2", "full-text"=>"2", "pdf"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"9"}
  • {"unique-ip"=>"7", "full-text"=>"7", "pdf"=>"3", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2020", "month"=>"10"}

Relative Metric

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