Journal: The Journal of Biological Chemistry

Article Title: Identification of Ror2 as a Hypoxia-inducible Factor Target in von Hippel-Lindau-associated Renal Cell Carcinoma *

doi: 10.1074/jbc.M109.073924

Figure Lengend Snippet: Minimal hypoxia induction of Ror2 expression. A, Ror2 expression is minimally induced after 48 h of exposure to cobalt chloride. Normoxic (N) and CoCl2 (C)-treated protein samples were immunoblotted with HIF-2α and Egln3 antibodies to show that HIF-2α and the HIF target Egln3 were induced in VHL(+) cell lines. Ror2 levels remained stable to this manipulation. Ku80 antibody was used as a loading control (LC). B, left, ROR2 mRNA levels are minimally induced under hypoxic-like conditions in RCC4 cells. qRT-PCR analysis of the hypoxia mimetic cobalt chloride (CoCl2) transcriptional induction after 24 h in the VHL expressing cell line RCC4 3-14 demonstrate induction of the HIF target gene EGLN3 (**, p = 0.0015) in response to treatment with hypoxia mimetic, confirming HIF transcriptional activity. Ror2 mRNA was not significantly induced under these conditions. Right, Ror2 mRNA levels are minimally induced under hypoxic-like conditions in 786-0 WT8 cells. qRT-PCR analysis of the hypoxia mimetic DMOG transcriptional induction after 24 h in the VHL expressing cell line 786-0 WT8 demonstrate induction of HIF target genes EGLN3 (**, p = 0.0091) and GLUT1 (**, p = 0.0015) in response to treatment with the hypoxia mimetic, confirming HIF transcriptional activity. Ror2 transcript levels show minimal enrichment upon treatment. Transcript values are shown as normalized to the β-actin RNA internal standard and relative to the unstimulated cells of each set of paired cells. Error bars represent ± S.E.

Article Snippet: The HIF-1α antibody was obtained from BD Transductions (Franklin Lakes, NJ) and the Glut1 and Egln3 antibodies were obtained from Novus Biologicals (Littleton, CO).

Techniques: Expressing, Quantitative RT-PCR, Activity Assay

Journal: Oncology reports

Article Title: Glucose-dependent glucose transporter 1 expression and its impact on viability of thyroid cancer cells.

doi: 10.3892/or.2014.3673

Figure Lengend Snippet: Figure 2. Effect of hypoglycemia, normoglycemia and hyperglycemia on the level of GLUT1 in plasma membrane in normoxic and hypoxic conditions in FTC-133 and 8305c cells. GLUT1, glucose transporter 1.

Article Snippet: The antibodies from Correspondence to: dr anna krześlak, department of Cytobiochemistry, Faculty of Biology and environmental Protection, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland E-mail: krzeslak@biol.uni.lodz.pl Key words: GLUT1, HIF1α, AKT1, glucose uptake, cell viability santa Cruz Biotechnology, Inc., (santa Cruz, Ca, Usa) used were: mouse monoclonal anti-GLUT3, rabbit polyclonal anti-HIF1α, mouse monoclonal anti-β-actin, goat polyclonal anti-rabbit IgG-HRP and goat polyclonal anti-mouse IgG-HRP.

Techniques: Clinical Proteomics, Membrane

Journal: Oncology reports

Article Title: Glucose-dependent glucose transporter 1 expression and its impact on viability of thyroid cancer cells.

doi: 10.3892/or.2014.3673

Figure Lengend Snippet: Figure 1. Effect of hypoglycemia, normoglycemia and hyperglycemia on the expression of GLUT1, GLUT3 and HIF1α in normoxic and hypoxic condi- tions in FTC-133 (A) and 8305c cells (B). Differences in GLUT1 and HIF1α expression levels in FTC-133 and 8305c cells (C). The immunodetection of GLUT1 and HIF1α in lysates from the cells treated with CoCl2 was performed on one blot to show the expression differences. GLUT1, glucose transporter 1; HIF1α, hypoxia inducible factor α.

Article Snippet: The antibodies from Correspondence to: dr anna krześlak, department of Cytobiochemistry, Faculty of Biology and environmental Protection, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland E-mail: krzeslak@biol.uni.lodz.pl Key words: GLUT1, HIF1α, AKT1, glucose uptake, cell viability santa Cruz Biotechnology, Inc., (santa Cruz, Ca, Usa) used were: mouse monoclonal anti-GLUT3, rabbit polyclonal anti-HIF1α, mouse monoclonal anti-β-actin, goat polyclonal anti-rabbit IgG-HRP and goat polyclonal anti-mouse IgG-HRP.

Techniques: Expressing, Immunodetection

Journal: Oncology reports

Article Title: Glucose-dependent glucose transporter 1 expression and its impact on viability of thyroid cancer cells.

doi: 10.3892/or.2014.3673

Figure Lengend Snippet: Figure 3. (A) Reduced expression of GLUT1 in FTC-133 and 8305c cells growing in hyperglycemia is correlated with reduced phosphorylation of Ser473 AKT1. (B) The intensity of the bands was analyzed by densitometry. GLUT1, glucose transporter 1.

Article Snippet: The antibodies from Correspondence to: dr anna krześlak, department of Cytobiochemistry, Faculty of Biology and environmental Protection, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland E-mail: krzeslak@biol.uni.lodz.pl Key words: GLUT1, HIF1α, AKT1, glucose uptake, cell viability santa Cruz Biotechnology, Inc., (santa Cruz, Ca, Usa) used were: mouse monoclonal anti-GLUT3, rabbit polyclonal anti-HIF1α, mouse monoclonal anti-β-actin, goat polyclonal anti-rabbit IgG-HRP and goat polyclonal anti-mouse IgG-HRP.

Techniques: Expressing, Phospho-proteomics

Journal: Oncology reports

Article Title: Glucose-dependent glucose transporter 1 expression and its impact on viability of thyroid cancer cells.

doi: 10.3892/or.2014.3673

Figure Lengend Snippet: Figure 6. (A) GLUT1 mRNA, (B) total protein and (C) membrane expression levels in FTC-133 and 8305c cells 48 or 72 h after siRNA treatment. GLUT1, glucose transporter 1.

Article Snippet: The antibodies from Correspondence to: dr anna krześlak, department of Cytobiochemistry, Faculty of Biology and environmental Protection, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland E-mail: krzeslak@biol.uni.lodz.pl Key words: GLUT1, HIF1α, AKT1, glucose uptake, cell viability santa Cruz Biotechnology, Inc., (santa Cruz, Ca, Usa) used were: mouse monoclonal anti-GLUT3, rabbit polyclonal anti-HIF1α, mouse monoclonal anti-β-actin, goat polyclonal anti-rabbit IgG-HRP and goat polyclonal anti-mouse IgG-HRP.

Techniques: Membrane, Expressing

Journal: Oncology reports

Article Title: Glucose-dependent glucose transporter 1 expression and its impact on viability of thyroid cancer cells.

doi: 10.3892/or.2014.3673

Figure Lengend Snippet: Figure 7. Effect of GLUT1 expression downregulation on (A) FTC-133 and (B) 8305c cell viability. Data are presented as the average of at least three independent experiments performed in triplicate (± SD) *P<0.05, **P<0.01 and ***P<0.001. GLUT1, glucose transporter 1.

Article Snippet: The antibodies from Correspondence to: dr anna krześlak, department of Cytobiochemistry, Faculty of Biology and environmental Protection, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland E-mail: krzeslak@biol.uni.lodz.pl Key words: GLUT1, HIF1α, AKT1, glucose uptake, cell viability santa Cruz Biotechnology, Inc., (santa Cruz, Ca, Usa) used were: mouse monoclonal anti-GLUT3, rabbit polyclonal anti-HIF1α, mouse monoclonal anti-β-actin, goat polyclonal anti-rabbit IgG-HRP and goat polyclonal anti-mouse IgG-HRP.

Techniques: Expressing

Journal: Oncology reports

Article Title: Glucose-dependent glucose transporter 1 expression and its impact on viability of thyroid cancer cells.

doi: 10.3892/or.2014.3673

Figure Lengend Snippet: Figure 8. Effect of GLUT1 expression downregulation on glucose analog 2-NBDG uptake to (A) FTC-133 and (B) 8305c cells. Data are presented as the average of at least three independent experiments performed in tripli- cate (± SD) *P<0.05, **P<0.01 and ***P<0.001. GLUT1, glucose transporter 1.

Article Snippet: The antibodies from Correspondence to: dr anna krześlak, department of Cytobiochemistry, Faculty of Biology and environmental Protection, University of Lodz, Pomorska 141/143, 90-236 Lodz, Poland E-mail: krzeslak@biol.uni.lodz.pl Key words: GLUT1, HIF1α, AKT1, glucose uptake, cell viability santa Cruz Biotechnology, Inc., (santa Cruz, Ca, Usa) used were: mouse monoclonal anti-GLUT3, rabbit polyclonal anti-HIF1α, mouse monoclonal anti-β-actin, goat polyclonal anti-rabbit IgG-HRP and goat polyclonal anti-mouse IgG-HRP.

Techniques: Expressing

Journal: The Journal of Experimental Medicine

Article Title: HDL and Glut1 inhibition reverse a hypermetabolic state in mouse models of myeloproliferative disorders

doi: 10.1084/jem.20121357

Figure Lengend Snippet: Enhanced glycolytic activity in Abca1 −/− Abcg1 −/− leukocytes. (A) Ex vivo characterization of the glucose binding and/or uptake in WT and Abca1 −/− Abcg1 −/− BM leukocyte subpopulations (i.e., CD115 + monocytes, CD115 − GrI + neutrophils, and TCRb + lymphocytes) using a fluorescent d -glucose analogue (2-NBDG). (B) Glucose uptake normalized by the amount of BM leukocytes. (C) mRNA expression of glucose transporters (Gluts) in freshly isolated WT and Abca1 −/− Abcg1 −/− BM cells. (D) Cell surface expression of Glut1 was also quantified in WT and Abca1 −/− Abcg1 −/− BM neutrophils, monocytes, and lymphocytes. (E) Mitochondrial membrane potential measured by fluorescent TMRE dye. All results are means ± SEM and are representative of two independent experiments ( n = 5–6 animals per groups). *, P < 0.05 versus controls. MFI, mean fluorescence intensity. (F) Glut1 expression after BM cells were treated for 72 h with the indicated growth factors and in the presence or absence of 1 µM farnesyl transferase inhibitor (FTI) or 10 µM PI3K inhibitor (LY294002). Values were normalized to ribosomal 18S. (G) Effect of IL-3 treatment and cholesterol depletion by cyclodextrin (CD) on [ 14 C]glucose conversion into CO 2 in WT and Abca1 −/− Abcg1 −/− BM cells. (H) mRNA expression of Hk2, PFK isoform p (PFKp), ACL, fumarase, and SDH subunit b (SDHb) in WT and Abca1 −/− Abcg1 −/− BM-derived cells untreated or treated for 72 h with IL-3. Values were normalized to ribosomal 18S. Results are means ± SEM of cultures from three independent mice. *, P < 0.05 versus WT controls; § , P < 0.05 versus untreated condition.

Article Snippet: In independent experiments, premade control and Glut1 shRNA lentiviral particles (Santa Cruz Biotechnology, Inc.) were used to transduce WT and Abca1 −/− Abcg1 −/− BM cells.

Techniques: Activity Assay, Ex Vivo, Binding Assay, Expressing, Isolation, Membrane, Fluorescence, Derivative Assay

Journal: The Journal of Experimental Medicine

Article Title: HDL and Glut1 inhibition reverse a hypermetabolic state in mouse models of myeloproliferative disorders

doi: 10.1084/jem.20121357

Figure Lengend Snippet: Inhibition of Glut1 or overexpression of the human apoA-I transgene rescues adipose atrophy in Abca1 −/− Abcg1 −/− BM chimeras. (A) Effect of IL-3 treatment on [ 14 C]glucose conversion into lipids in WT and Abca1 −/− Abcg1 −/− BM cells in the presence or absence of 50 µM fasentin, a Glut1 inhibitor. (B) Effect of Fasentin on IL-3–mediated proliferation in WT and Abca1 −/− Abcg1 −/− BM cells. (C and D) Effect of Fasentin on LPS-mediated inflammatory (C) and glycolytic (D) responses in WT and Abca1 −/− Abcg1 −/− BM-derived macrophages. Values were normalized to ribosomal 18S. Results are means ± SEM of three independent experiments performed in triplicate. *, P < 0.05 versus WT mice; § , P < 0.05 versus LPS treatment. (E and F) Quantification of Glut1 cell surface expression (E) and 2-NBDG uptake by flow cytometry (F) in CD45 + leukocytes isolated from the BM of WT recipients mice transplanted with WT or Abca1 −/− Abcg1 −/− BM transduced with a lentivirus encoding Glut1 shRNA 7 wk after reconstitution. (G and H) Peripheral leukocyte counts (G) and epididymal fat mass (H) of these mice at the end of the experiment. (I and J) Histograms showing epididymal fat mass loss (I) and Glut1 cell surface expression (J) in CD45 + peripheral leukocytes in chow-fed transgenic recipient mice overexpressing the human apoA-I transgene transplanted with Abca1 −/− Abcg1 −/− BM. Results are ± SEM of an experiment of four to six animals per group. *, P < 0.05 versus WT recipients transplanted with WT BM; § , P < 0.05 versus control retrovirus. MFI, mean fluorescence intensity.

Article Snippet: In independent experiments, premade control and Glut1 shRNA lentiviral particles (Santa Cruz Biotechnology, Inc.) were used to transduce WT and Abca1 −/− Abcg1 −/− BM cells.

Techniques: Inhibition, Over Expression, Derivative Assay, Expressing, Flow Cytometry, Isolation, Transduction, shRNA, Transgenic Assay, Control, Fluorescence

Journal: The Journal of Experimental Medicine

Article Title: HDL and Glut1 inhibition reverse a hypermetabolic state in mouse models of myeloproliferative disorders

doi: 10.1084/jem.20121357

Figure Lengend Snippet: HDL prevents the adipose tissue atrophy and enhanced glucose oxidation caused by Mpl-W515L and Flt3-ITD activating mutations. (A) Representative dot plots and quantification of the splenic myeloid cells (CD115 + monocytes and CD115 − Gr1 + neutrophils) of WT and ApoA-I transgenic recipient mice transplanted with Mpl-W515L– and Flt3-ITD–transduced BM cells. (B and C) Quantification of the myeloid cells (B) and epididymal fat mass (C) of these mice. (D, E, F, H, and I) RQ measured by indirect calorimetry (D, E, H, and I) and quantification of the nocturnal glucose oxidation of these mice (F). (G) Quantification of the cell surface expression of Glut1 in CD45 + leukocytes of these mice. Results are mean ± SEM of an experiment of four to five animals per group. *, P < 0.05 versus control mice; § , P < 0.05 versus respective WT recipients. MFI, mean fluorescence intensity.

Article Snippet: In independent experiments, premade control and Glut1 shRNA lentiviral particles (Santa Cruz Biotechnology, Inc.) were used to transduce WT and Abca1 −/− Abcg1 −/− BM cells.

Techniques: Transgenic Assay, Expressing, Control, Fluorescence

Journal: Frontiers in oncology

Article Title: Long Non-Coding RNA TMPO-AS1 Promotes GLUT1-Mediated Glycolysis and Paclitaxel Resistance in Endometrial Cancer Cells by Interacting With miR-140 and miR-143.

doi: 10.3389/fonc.2022.912935

Figure Lengend Snippet: FIGURE 1 | GLUT1 Expression and Clinical Importance in EC. (A) The mRNA expression of GLUT1 (cancer versus normal tissues) in pan-cancers was analyzed with the Oncomine database. The graphic demonstrates the number of datasets that meet our threshold for each cancer type. Red: up-regulation; blue: down- regulation. (B) The heatmap was generated by the ImaGEO database using two EC data sets (GSE17025 and GSE56026). (C) Up- or down-regulated genes in EC tissues compared to normal tissues (ImaGEO). (D) The GEO dataset was analyzed for GLUT1 expression in stage I EC samples and normal endometrium samples. (E) The expression of GLUT1 protein was examined in EC tissue and adjacent normal tissues (Human Protein Atlas database; scale bar: 100 mm). (F) The probability of overall survival in EC patients expressing high or low GLUT1 levels was assessed using the KM plotter database.

Article Snippet: May 2022 | Volume 12 | Article 912935 GLUT1 silencing was conducted in the Ishikawa cell line using shRNA targeting GLUT1 (Santa Cruz Biotechnologies, Santa Cruz, CA) or control shRNA (Santa Cruz Biotechnologies) and Lipofectamine 3000 reagent.

Techniques: Expressing, Generated

Journal: Frontiers in oncology

Article Title: Long Non-Coding RNA TMPO-AS1 Promotes GLUT1-Mediated Glycolysis and Paclitaxel Resistance in Endometrial Cancer Cells by Interacting With miR-140 and miR-143.

doi: 10.3389/fonc.2022.912935

Figure Lengend Snippet: FIGURE 2 | Critical Roles of GLUT1 in Promoting EC Cell Phenotypes. (A) Western blotting analysis of GLUT1 protein expression in a normal endometrial cell line (EM) and human EC cell lines. (B) Western blotting analysis of GLUT1 expression in GLUT1-overexpressing HHUA cells and Ishikawa cells with GLUT1 silencing. (C–F) EC cell proliferation (C), invasion (D), glucose consumption (E), and lactate production (F) assays following GLUT1 overexpression or knockdown. (G, H) GLUT1-overexpressing HHUA cells (G) and Ishikawa cells after knockdown of GLUT1 (H) were treated with different concentrations of paclitaxel, and cell viability was examined using the CCK-8 assay. Vec: vector. ***P < 0.001.

Article Snippet: May 2022 | Volume 12 | Article 912935 GLUT1 silencing was conducted in the Ishikawa cell line using shRNA targeting GLUT1 (Santa Cruz Biotechnologies, Santa Cruz, CA) or control shRNA (Santa Cruz Biotechnologies) and Lipofectamine 3000 reagent.

Techniques: Western Blot, Expressing, Over Expression, Knockdown, CCK-8 Assay, Plasmid Preparation

Journal: Frontiers in oncology

Article Title: Long Non-Coding RNA TMPO-AS1 Promotes GLUT1-Mediated Glycolysis and Paclitaxel Resistance in Endometrial Cancer Cells by Interacting With miR-140 and miR-143.

doi: 10.3389/fonc.2022.912935

Figure Lengend Snippet: FIGURE 3 | KEGG Pathway and Wiki Pathway Enrichment Analysis of GLUT1-Associated Genes in EC. (A) Genes that were found to be highly co-expressed with GLUT1 in the TCGA EC dataset from the LinkedOmics database were chosen. (B, C) Gene set enrichment analysis for positively correlated genes of GLUT1 in TCGA EC tissues. Enrichment analysis for KEGG pathways (B) and Wikipathway cancer (C) was performed using the LinkedOmics database.

Article Snippet: May 2022 | Volume 12 | Article 912935 GLUT1 silencing was conducted in the Ishikawa cell line using shRNA targeting GLUT1 (Santa Cruz Biotechnologies, Santa Cruz, CA) or control shRNA (Santa Cruz Biotechnologies) and Lipofectamine 3000 reagent.

Techniques:

Journal: Frontiers in oncology

Article Title: Long Non-Coding RNA TMPO-AS1 Promotes GLUT1-Mediated Glycolysis and Paclitaxel Resistance in Endometrial Cancer Cells by Interacting With miR-140 and miR-143.

doi: 10.3389/fonc.2022.912935

Figure Lengend Snippet: FIGURE 4 | GLUT1 is a Positive Upstream Regulator of MMP1, MMP14, and Cyclin D1. (A) The expression of GLUT1 was positively correlated with the expression of MMP1, MMP14, and Cyclin D1 in EC tissues from the TCGA dataset. (B) qRT-PCR analysis of the indicated genes in EC cells after overexpression or knockdown of GLUT1. (C) The expression of MMP1, MMP14, and Cyclin D1 in TCGA EC and normal tissues (ENCORI database). ***P < 0.001.

Article Snippet: May 2022 | Volume 12 | Article 912935 GLUT1 silencing was conducted in the Ishikawa cell line using shRNA targeting GLUT1 (Santa Cruz Biotechnologies, Santa Cruz, CA) or control shRNA (Santa Cruz Biotechnologies) and Lipofectamine 3000 reagent.

Techniques: Expressing, Quantitative RT-PCR, Over Expression, Knockdown

Journal: Frontiers in oncology

Article Title: Long Non-Coding RNA TMPO-AS1 Promotes GLUT1-Mediated Glycolysis and Paclitaxel Resistance in Endometrial Cancer Cells by Interacting With miR-140 and miR-143.

doi: 10.3389/fonc.2022.912935

Figure Lengend Snippet: FIGURE 5 | GLUT1 is a Target Gene of MiR-140 and MiR-143. (A) Venn diagram showing the overlapping miRNAs predicted by three online databases. (B) Computational prediction of duplex formation between miR-140/miR-143 and the GLUT1 3′-UTR region. (C) The expression of miR-140/miR-143 in TCGA EC tissues and normal tissues (miR-TV database). (D) qRT-PCR analysis of miR-140/miR-143 in EC and EM cells. (E) GLUT1 protein expression was examined using western blotting assays in EC cells transfected as indicated. (F) Luciferase reporter assays in Ishikawa cells transfected with the wild-type (WT) or mutant (MUT) GLUT1 3′-UTR, as well as miR-140/miR-143 mimic or a control mimic. ***P < 0.001.

Article Snippet: May 2022 | Volume 12 | Article 912935 GLUT1 silencing was conducted in the Ishikawa cell line using shRNA targeting GLUT1 (Santa Cruz Biotechnologies, Santa Cruz, CA) or control shRNA (Santa Cruz Biotechnologies) and Lipofectamine 3000 reagent.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transfection, Luciferase, Mutagenesis, Control

Journal: Frontiers in oncology

Article Title: Long Non-Coding RNA TMPO-AS1 Promotes GLUT1-Mediated Glycolysis and Paclitaxel Resistance in Endometrial Cancer Cells by Interacting With miR-140 and miR-143.

doi: 10.3389/fonc.2022.912935

Figure Lengend Snippet: FIGURE 8 | TMPO-AS1 Induces GLUT1 Expression by Repressing MiR-140 and MiR-143 Levels. (A) GLUT1 protein expression was measured in Ishikawa and HHUA cells transfected with (or without) TMPO-AS1 siRNA, along with (or without) miR-140/miR-143 inhibitor. (B) Examination of MMP1, MMP14, and Cyclin D1 expression in Ishikawa cells transfected as indicated. (C) Correlation analysis between TMPO-AS1 and GLUT1 or between TMPO-AS1 and miR-140/miR-143. ***P < 0.001.

Article Snippet: May 2022 | Volume 12 | Article 912935 GLUT1 silencing was conducted in the Ishikawa cell line using shRNA targeting GLUT1 (Santa Cruz Biotechnologies, Santa Cruz, CA) or control shRNA (Santa Cruz Biotechnologies) and Lipofectamine 3000 reagent.

Techniques: Expressing, Transfection

Journal: Frontiers in oncology

Article Title: Long Non-Coding RNA TMPO-AS1 Promotes GLUT1-Mediated Glycolysis and Paclitaxel Resistance in Endometrial Cancer Cells by Interacting With miR-140 and miR-143.

doi: 10.3389/fonc.2022.912935

Figure Lengend Snippet: FIGURE 10 | Graphical Abstract showing that the LncRNA TMPO-AS1 Regulates Glycolysis and Paclitaxel Resistance in EC Cells by Affecting the MiR-140/MiR- 143-GLUT1 Pathway.

Article Snippet: May 2022 | Volume 12 | Article 912935 GLUT1 silencing was conducted in the Ishikawa cell line using shRNA targeting GLUT1 (Santa Cruz Biotechnologies, Santa Cruz, CA) or control shRNA (Santa Cruz Biotechnologies) and Lipofectamine 3000 reagent.

Techniques: