Journal: Cell Reports Medicine

Article Title: Exercise-induced crosstalk between immune cells and adipocytes in humans: Role of oncostatin-M

doi: 10.1016/j.xcrm.2023.101348

Figure Lengend Snippet:

Article Snippet: GDF15 , Bio-Techne LTD , 9279-GD-050.

Techniques: In Vitro, Recombinant, Plasmid Preparation, Fluorescence, Blocking Assay, Protease Inhibitor, Enzyme-linked Immunosorbent Assay, Bicinchoninic Acid Protein Assay, Multiplex Assay, Software, Real-time Polymerase Chain Reaction, Microscopy

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Growth differentiation factor 15 (GDF15) expression is elevated in gastric cancer and high serum GDF15 level is a poor prognostic factor. (A) GDF15 gene expression level in gastric cancer patients illustrated with boxplots by the Gene Expression Profiling Interactive Analysis (GEPIA) online database. (B) The Kaplan–Meier plotter online database was used to analyze the clinical effect of GDF15 gene expression in gastric cancer patients ( http://kmplot.com/analysis/ ). (C) Clinical effect of GDF15 serum levels (≥upper quartile 1066.79 ng/mL vs.

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Expressing, Gene Expression

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Clinical characteristics of patients with gastric cancer with different growth differentiation factor 15 (GDF15) expression of tumor tissues

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Expressing

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Clinical characteristics of patients with gastric cancer with low or high serum growth differentiation factor 15 (GDF15) levels

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques:

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Growth differentiation factor 15 (GDF15) expression is essential for cell proliferation and migration of gastric cancer cells. (A, C) siGDF15 (180 pmol for 3 × 10 5 cells in a 6‐cm dish for 48 h) or (B, D) pcDNA‐GDF15 (pGDF15, 6 μg for 3 × 10 5 cells in a 6‐cm dish for 12 h, followed by replacement of fresh medium for a total of 48 h) were used to knockdown or overexpress GDF15. Efficiencies of knockdown and overexpression were analyzed by quantitative real‐time PCR. (A, B) After transfection with siGDF15 or pGDF15, cells were reseeded with a density of 3000 cells per well in a 96‐well plate. Cell proliferation was analyzed with sulforhodamine B (SRB) assay. (C, D) After transfection with siGDF15 or pGDF15, cells were reseeded with a density of 1 × 10 5 cells per Transwell insert. Cell migration was determined by Transwell migration assay (siGDF15: AGS, NUGC‐3, and TSGH9201 for 12, 16, and 24 h migration, respectively; magnification, 200×) (pGDF15: AGS, NUGC‐3, and TSGH9201 for 8, 12, and 24 h migration, respectively; magnification, 100×). Graph is presented as mean ± SEM ( n ≥ 3). *Significant vs. individual control.

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Expressing, Migration, Knockdown, Over Expression, Real-time Polymerase Chain Reaction, Transfection, Sulforhodamine B Assay, Transwell Migration Assay, Control

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Growth differentiation factor 15 (GDF15) contributes to cisplatin resistance in human gastric cancer cells. (A–C) GDF15 (A) gene and (B) protein expressions and (C) released GDF15 level between parental (P) and cisplatin‐resistant (CisR) gastric cancer cells were analyzed with quantitative real‐time PCR, western blotting, and ELISA assays, respectively. (D, E) Cisplatin sensitivity (48 h) of the gastric cancer cells was assessed using (D) sulforhodamine B (SRB) assay and (E) propidium iodide (PI) exclusion assay. (F–H) Effects of (F) GDF15 neutralizing Ab (GDF15 NAb), (G) recombinant human GDF15 (rhGDF15), and (H) GDF15 overexpression on sensitivity of cisplatin were evaluated with SRB assay. G, GDF15 plasmid; V, empty vector. Quantitative real‐time PCR and western blotting were used to validate the efficiencies of GDF15 knockdown or overexpression, respectively. Graph is presented by mean ± SEM ( n ≥ 3). *Significant vs. individual control. ** , ***Significant, rhGDF15 (20 and 50 ng/mL) vs. individual control.

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay, Sulforhodamine B Assay, Exclusion Assay, Recombinant, Over Expression, Plasmid Preparation, Knockdown, Control

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Growth differentiation factor 15 (GDF15)‐upregulated xCT expression through the eukaryotic initiation factor 2α (eIF2α)‐activating transcription factor 4 (ATF4) pathway enhances intracellular glutathione (GSH) levels in cisplatin‐resistant gastric cancer cells. (A) Knockdown efficiency was validated using western blotting. (B) Effects of siGDF15 and glial cell‐derived neurotrophic factor family receptor a‐like siRNA (siGFRAL) on GSH levels were evaluated using the GSH detection kit. (C, D) After treatment of GDF15‐knockdown cisplatin‐resistant (CisR) cells with cisplatin (24 h), intracellular and mitochondrial reactive oxygen species were evaluated with (C) dichlorodihydro‐fluorescein (DCF) and (D) MitoSox Red using flow cytometry. (E) GDF15 and xCT gene expressions were evaluated using quantitative real‐time PCR. (F) Protein expression of GDF15 and the eIF2α‐xCT pathway were evaluated using western blotting. (G) After transfections with different xCT promoters (WT, antioxidant‐responsive element [ARE]‐mutant, and amino acid response element [AARE]‐mutant), the cells were further transfected with siGDF15. Graph is presented as mean ± SEM ( n ≥ 3). *Significant vs. individual control. **Significant vs. WT/ARE‐mutant‐xCT promoters.

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Expressing, Knockdown, Western Blot, Derivative Assay, Flow Cytometry, Real-time Polymerase Chain Reaction, Transfection, Mutagenesis, Control

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Eukaryotic initiation factor 2α (eIF2α)‐activating transcription factor 4 (ATF4)‐xCT‐elevated glutathione (GSH) contributes to growth differentiation factor 15 (GDF15)‐mediated cisplatin resistance in gastric cancer cells. (A, B, D) Cells were transfected with pcDNA‐GDF15. G, GDF15 plasmid; V, empty vector. (A, D) Gene expressions of GDF15 and xCT were evaluated using quantitative real‐time PCR. (B) The eIF2α‐ATF4‐xCT pathway was evaluated using western blotting. (C) After transfection with different types of xCT promoters, HEK293T cells were further transfected with pcDNA‐GDF15. (D) After overexpression of GDF15, the effects of sulfasalazine (SSA, 350 μM) and buthionine sulfoximine (BSO, 0.5 mM) on cisplatin sensitivity (48 h) were evaluated with propidium iodide (PI) exclusion assay. Graph is presented as mean ± SEM ( n ≥ 3). *Significant vs. individual control. **Significant vs. WT/antioxidant‐responsive element (ARE)‐mutant xCT promoters. # Significant vs. pcDNA. ## Significant vs. cisplatin treatment. AARE, amino acid response element; Con, control.

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Transfection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Western Blot, Over Expression, Exclusion Assay, Control, Mutagenesis

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Growth differentiation factor 15 (GDF15)/ glial cell‐derived neurotrophic factor family receptor a‐like (GFRAL)‐mediated signaling in cisplatin‐resistant gastric cancer cells could be through general control nonderepressible 2 (GCN2). (A, B) siGDF15 and siGFRAL were transfected into (A) AGS cisplatin‐resistant (CisR) and (B) NUGC‐3CisR cells. (C) AGSCisR and (D) NUGC‐3CisR cells were treated with SPP86 (5 μΜ) for 24 h. Upstream regulators of the eukaryotic initiation factor 2α (eIF2α) and eIF2α‐activating transcription factor 4 (ATF4)‐xCT pathways were analyzed using western blotting. Graph is presented as mean ± SEM ( n ≥ 3). *Significant vs. individual control (Con). PERK, PKR‐like endoplasmic reticulum kinase; PKR, protein kinase R.

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Derivative Assay, Control, Transfection, Western Blot

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: General control nonderepressible 2 (GCN2) is responsible for growth differentiation factor 15 (GDF15)‐mediated glial cell‐derived neurotrophic factor family receptor a‐like (GFRAL)‐eukaryotic initiation factor 2α (eIF2α)‐activating transcription factor 4 (ATF4)‐xCT signaling and cisplatin resistance. (A, B, D) After GDF15 overexpression by pcDNA‐GDF15 (G, pcDNA‐GDF15; V, pcDNA alone), cells were treated with siRNAs against (A) protein kinase R (PKR), (B) heme‐regulated eIF2α kinase (HRI), and (D) GCN2 for 48 h. The effect of siRNAs against PKR, HRI, and GCN2 on GDF15‐mediated eIF2α‐ATF4‐xCT regulation was assessed using western blotting. (C) Effects of siHRI and siPKR on cisplatin resistance in cisplatin‐resistant (CisR) cells was evaluated with sulforhodamine B (SRB) assay. Graph is presented as mean ± SEM ( n ≥ 3). *Significant vs. individual control. **Significant vs. siScr with GDF15 overexpression.

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Control, Derivative Assay, Over Expression, Western Blot, Sulforhodamine B Assay

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Proposed mechanism of growth differentiation factor 15 (GDF15)‐mediated cisplatin resistance. In the present study, we found that GDF15‐elevated glutathione (GSH) through the glial cell‐derived neurotrophic factor family receptor a‐like (GFRAL)‐general control nonderepressible 2 (GCN2)‐eukaryotic initiation factor 2α (eIF2α)‐activating transcription factor 4 (ATF4)‐xCT pathway enhances cisplatin resistance for gastric cancer. Figure was created by Servier Medical Art. PKR, protein kinase R; RET, rearranged during transfection; ROS, reactive oxygen species.

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Derivative Assay, Control, Transfection