Arsenite-Induced Pseudo-Hypoxia Results in Loss of Anchorage-Dependent Growth in BEAS-2B Pulmonary Epithelial Cells
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{"title"=>"Arsenite-induced pseudo-hypoxia results in loss of anchorage-dependent growth in BEAS-2B pulmonary epithelial cells", "type"=>"journal", "authors"=>[{"first_name"=>"Fei", "last_name"=>"Zhao", "scopus_author_id"=>"55348371600"}, {"first_name"=>"Scott W.", "last_name"=>"Malm", "scopus_author_id"=>"56448783900"}, {"first_name"=>"Alyssa N.", "last_name"=>"Hinchman", "scopus_author_id"=>"56449231000"}, {"first_name"=>"Hui", "last_name"=>"Li", "scopus_author_id"=>"57060900900"}, {"first_name"=>"Connor G.", "last_name"=>"Beeks", "scopus_author_id"=>"56449273400"}, {"first_name"=>"Walter T.", "last_name"=>"Klimecki", "scopus_author_id"=>"35476853200"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"600783728", "sgr"=>"84918556503", "issn"=>"19326203", "pmid"=>"25513814", "scopus"=>"2-s2.0-84918556503", "doi"=>"10.1371/journal.pone.0114549", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)"}, "id"=>"e8f27e34-245e-3776-9eb1-a8d103e24467", "abstract"=>"Epidemiology studies have established a strong link between lung cancer and arsenic exposure. Currently, the role of disturbed cellular energy metabolism in carcinogenesis is a focus of scientific interest. Hypoxia inducible factor-1 alpha (HIF-1A) is a key regulator of energy metabolism, and it has been found to accumulate during arsenite exposure under oxygen-replete conditions. We modeled arsenic-exposed human pulmonary epithelial cells in vitro with BEAS-2B, a non-malignant lung epithelial cell line. Constant exposure to 1 microM arsenite (As) resulted in the early loss of anchorage-dependent growth, measured by soft agar colony formation, beginning at 6 weeks of exposure. This arsenite exposure resulted in HIF-1A accumulation and increased glycolysis, similar to the physiologic response to hypoxia, but in this case under oxygen-replete conditions. This \"pseudo-hypoxia\" response was necessary for the maximal acquisition of anchorage-independent growth in arsenite-exposed BEAS-2B. The HIF-1A accumulation and induction in glycolysis was sustained throughout a 52 week course of arsenite exposure in BEAS-2B. There was a time-dependent increase in anchorage-independent growth during the exposure to arsenite. When HIF-1A expression was stably suppressed, arsenite-induced glycolysis was abrogated, and the anchorage-independent growth was reduced. These findings establish that arsenite exerts a hypoxia-mimetic effect, which plays an important role in the subsequent gain of malignancy-associated phenotypes.", "link"=>"http://www.mendeley.com/research/arseniteinduced-pseudohypoxia-results-loss-anchoragedependent-growth-beas2b-pulmonary-epithelial-cel", "reader_count"=>8, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>2, "Student > Ph. D. Student"=>3, "Student > Postgraduate"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>2, "Student > Ph. D. Student"=>3, "Student > Postgraduate"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Environmental Science"=>1, "Agricultural and Biological Sciences"=>4, "Medicine and Dentistry"=>2}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>4}, "Unspecified"=>{"Unspecified"=>1}, "Environmental Science"=>{"Environmental Science"=>1}}, "reader_count_by_country"=>{"United States"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1844801"], "description"=>"<p>A) Immunoblot analysis of HIF-1A in BEAS-2B treated with 0–8 µM arsenite for 48 hours. B) Immunoblot analysis of HIF-1A in BEAS-2B treated with 1 µM arsenite for 0–48 hours. C) Immunoblot analysis of nuclear and cytosolic fractions of BEAS-2B, control or treated with 1 µM arsenite for 2 weeks, probed for HIF-1A, Lamin A (a nuclear marker) and tubulin (a cytosolic marker). D) Immunofluorescence staining of HIF-1A in BEAS-2B, control or treated with 1 µM arsenite for 2 weeks, arrows show HIF-1A nuclear accumulation. E) QPCR of HIF-1A mRNA in BEAS-2B treated with 1 µM arsenite for 0–4 weeks, bars represent mean, 1 standard deviation. F) Half-life measurement of HIF-1A in BEAS-2B, control or treated with 1 µM arsenite for 2 weeks, protein synthesis blocked with cycloheximide (CHX) for 0–10 min, followed by HIF-1A immunoblot. G) Quantification of HIF-1A protein half-life (t1/2). Densitometry of HIF-1A normalized to Tubulin was used for calculation. Points represent mean, +/− 1 standard deviation, 3 independent replicates. *p<0.05.</p>", "links"=>[], "tags"=>["agar colony formation", "glycolysi", "Hypoxia Inducible Factor", "52 week course", "arsenite exposure", "Energy metabolism", "hif", "beas", "1 µ M arsenite"], "article_id"=>1272352, "categories"=>["Uncategorised"], "users"=>["Fei Zhao", "Scott W. Malm", "Alyssa N. Hinchman", "Hui Li", "Connor G. Beeks", "Walter T. Klimecki"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0114549.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Arsenite_causes_HIF_1A_accumulation_translocation_in_BEAS_2B_/1272352", "title"=>"Arsenite causes HIF-1A accumulation/translocation in BEAS-2B.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-16 02:49:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/1844805"], "description"=>"<p>A) Immunoblot analysis of HIF-1A in BEAS-2B, vector control and transiently transfected with degradation-resistant HIF-1A mutant. B) Lactate levels (percent vector control) in cells described in 2A (Absolute lactate production in vector control: 0.729±0.054 µmol/10<sup>6</sup>cells/hr). Bars represent mean, 1 standard deviation, from 3 independent replicates. *p<0.05. C) Intracellular metabolite concentration (percent control BEAS-2B) of 1 µM arsenite-exposed (2 weeks) BEAS-2B cells. Bars represent mean, 1 standard deviation, from 4 experimental replicates. For each metabolite, levels in arsenite-exposed BEAS-2B are significantly different compared to control (p<0.05).</p>", "links"=>[], "tags"=>["agar colony formation", "glycolysi", "Hypoxia Inducible Factor", "52 week course", "arsenite exposure", "Energy metabolism", "hif", "beas", "1 µ M arsenite"], "article_id"=>1272356, "categories"=>["Uncategorised"], "users"=>["Fei Zhao", "Scott W. Malm", "Alyssa N. Hinchman", "Hui Li", "Connor G. Beeks", "Walter T. Klimecki"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0114549.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Glycolysis_induction_by_HIF_1A_overexpression_in_BEAS_2B_/1272356", "title"=>"Glycolysis induction by HIF-1A overexpression in BEAS-2B.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-16 02:49:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/1844810"], "description"=>"<p>A) Representative images of soft agar growth over the course of 52 weeks of constant arsenite (1 µM) exposure. B) Colony counts in soft agar. Bars represent mean, 1 standard deviation, from 3 experimental replicates. C) Immunoblot analysis of HIF-1A and E-cadherin (E-cad) in BEAS-2B over the course of 52 weeks of constant arsenite (1 µM) exposure. D) Lactate levels (percent control) in BEAS-2B over the course of 52 weeks of constant arsenite (1 µM) exposure. Absolute lactate production in vector control: 0.733±0.017 µmol/10<sup>6</sup>cells/hr) Bars represent mean +1 standard deviation, from 3 experimental replicates. E) Percentage aneuploid cells in BEAS-2B treated with 1 µM arsenite for 0–52 weeks. Bars represent mean, +1 standard deviation, from 3 experimental replicates. *p<0.05.</p>", "links"=>[], "tags"=>["agar colony formation", "glycolysi", "Hypoxia Inducible Factor", "52 week course", "arsenite exposure", "Energy metabolism", "hif", "beas", "1 µ M arsenite"], "article_id"=>1272361, "categories"=>["Uncategorised"], "users"=>["Fei Zhao", "Scott W. Malm", "Alyssa N. Hinchman", "Hui Li", "Connor G. Beeks", "Walter T. Klimecki"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0114549.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Arsenite_induced_phenotypic_changes_in_BEAS_2B_/1272361", "title"=>"Arsenite-induced phenotypic changes in BEAS-2B.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-16 02:49:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/1844815"], "description"=>"<p>A) Immunoblot analysis of HIF-1A knockdown in BEAS-2B, short immunoblot exposure shown for MG132-treated samples; long immunoblot exposure shown for MG132-untreated samples. B) QPCR for HIF-1A mRNA. Bars represent mean, +1 standard deviation, from 5 experimental replicates. C) Lactate levels (percent control) in arsenite-exposed (denoted “As”, exposed for 8 weeks) and unexposed control (denoted “Ct”) BEAS-2B stably transfected with scrambled control shRNA (denoted “Vector”) or with shRNA targeting HIF1A (denoted “shHIF1A”) expression. Absolute lactate production in vector control: 0.696±0.04 µmol/10<sup>6</sup>cells/hr). Bars represent mean, +1 standard deviation, from 3 experimental replicates. D) Colony count of soft agar assay from BEAS-2B cells treated as described above in panel C. Bars represent mean, +1 standard deviation, from 3 experimental replicates. *p<0.05.</p>", "links"=>[], "tags"=>["agar colony formation", "glycolysi", "Hypoxia Inducible Factor", "52 week course", "arsenite exposure", "Energy metabolism", "hif", "beas", "1 µ M arsenite"], "article_id"=>1272366, "categories"=>["Uncategorised"], "users"=>["Fei Zhao", "Scott W. Malm", "Alyssa N. Hinchman", "Hui Li", "Connor G. Beeks", "Walter T. Klimecki"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0114549.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_suppressed_HIF_1A_expression_on_arsenite_mediated_transformation_/1272366", "title"=>"Effect of suppressed HIF-1A expression on arsenite mediated transformation.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-16 02:49:22"}

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Relative Metric

{"start_date"=>"2014-01-01T00:00:00Z", "end_date"=>"2014-12-31T00:00:00Z", "subject_areas"=>[]}
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