Journal: World Journal of Surgical Oncology

Article Title: LAIR-1 overexpression inhibits epithelial–mesenchymal transition in osteosarcoma via GLUT1-related energy metabolism

doi: 10.1186/s12957-020-01896-7

Figure Lengend Snippet: LAIR-1 inhibits Glut1-related glucose uptake in OS cells. a Heatmap showing the levels of differentially expressed mRNAs. b Top 20 KEGG pathway annotation categories for target gene functions of predicted mRNAs. c Selected significantly differentially expressed mRNA-related to EMT in RNA-seq data between two groups, *** P < 0.001. d qPCR validation of differentially expressed EMT-related genes in LV-NC and LV-LAIR-1-overexpressing OS cells, ** P < 0.01. e Glut1 expression analyzed by western blotting. f Immunofluorescence staining of Glut1 in the LV-LAIR-1-overexpressing OS cells. Scale bar = 50 μm. Data were obtained from at least two independent experiments.

Article Snippet: Total protein was extracted using a routine procedure and blotted with the following primary antibodies: LAIR-1 (sc-398141; Santa Cruz Biotechnology), phospho-Foxo1 (Ser256) (84192; Cell Signaling Technology, Danvers, MA, USA), Foxo1 (2880; Cell Signaling Technology), phospho-Akt (Ser473) (AF8355; Affinity Biosciences, Cincinnati, OH, USA), Akt (9272; Cell Signaling Technology), proliferating cell nuclear antigen (PCNA; BM0104; Boster Biotech Co., Ltd., Wuhan, China), Twist1 (ab50581; Abcam, Cambridge, UK), Glut1 (NB110-39113, Novus Biologicals, Littleton, CO, USA), and β-actin (30101ES50; Yeasen Biotech Co., Ltd., Shanghai, China).

Techniques: RNA Sequencing, Biomarker Discovery, Expressing, Western Blot, Immunofluorescence, Staining

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:

Journal: Diabetologia

Article Title: Low levels of glucose transporters and K+ATP channels in human pancreatic beta cells early in development.

doi: 10.1007/s00125-007-0644-x

Figure Lengend Snippet: Fig. 2 Beta cell production of GLUTs, GCK and Kþ ATP channels in human fetal pancreas development. Proportions of insulin-positive cells positive for GLUT2 (black bars), GLUT1 (sloping hatched bars), GCK (horizontally hatched bars), KIR6.2 (vertically hatched bars) and SUR1 (white bars) components of Kþ ATP channels were quantified by analysis of images prepared as in Fig. 1. Results are expressed as means±SEM of pancreases of 11–15 weeks (n=5), 16–19 weeks (n=5) and 20–36 weeks (n=6) of gestation, infant pancreas (3–7.5 months of age; n=3) and juvenile and adult pancreases (n=3). mo, months; w, weeks; y, year

Article Snippet: Sections were dewaxed and rehydrated, then incubated overnight with 1:100 dilutions of primary antibodies to GLUT1, GLUT2, GCK, KIR6.2 and SUR1 (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA).

Techniques:

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