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

Journal: International Journal of Oncology

Article Title: Effects of non-steroidal anti-inflammatory drug-activated gene-1 on Ganoderma lucidum polysaccharides-induced apoptosis of human prostate cancer PC-3 cells

doi: 10.3892/ijo.2018.4578

Figure Lengend Snippet: Expression levels of NAG-1, and its transcription factor EGR-1, are induced in PC-3 cells upon GLP treatment. (A) Quantification of the mRNA expression levels of NAG-1 and EGR-1 in a time-dependent manner (0-24 h) following treatment of PC-3 cells with 5 mg/ml GLP. Data are presented as the means ± standard error from three independent experiments. Two-way ANOVA with post hoc Bonferroni's correction for multiple comparison was used to determine statistical significance. * indicates time-dependent effects; # indicates effects of GLP treatment. * P<0.05 and ** P<0.01 compared with the 0 h group for each treatment. # P<0.05 and ## P<0.01 compared with the untreated control group at each time-point. (B) Quantification of the mRNA expression levels of NAG-1 and EGR-1 in a dose-dependent manner following treatment of PC-3 cells with GLP (0-10 mg/ml) for 24 h. Induction of EGR-1 and NAG-1 protein expression upon GLP treatment in a (C) time-dependent and (D) dose-dependent manner, as determined by western blotting. β-actin was used as an internal control. (E) GLP induced NAG-1 promoter activity, as determined by luciferase assay. Fold-changes were normalized to pRL-null expressing Renilla luciferase protein. (F) ELISA of the concentration of NAG-1 protein in the cell culture medium following treatment with 0-10 mg/ml GLP for 48 h. Data presented were normalized to the concentration of protein in lysates from each sample. All data are presented as the means ± standard error of three independent experiments. * P<0.05, ** P<0.01 compared with the control group (one-way ANOVA with Dunnett's correction). ANOVA, analysis of variance; EGR-1, early growth response-1; GLP, Ganoderma lucidum polysaccharides; NAG-1, non-steroidal anti-inflammatory drug-activated gene-1; RLU, relative light units.

Article Snippet: The Quantikine Human GDF15/NAG-1 ELISA kit (cat. no. DY957) was purchased from R&D Systems, Inc. (Minneapolis, MN, USA).

Techniques: Expressing, Comparison, Western Blot, Activity Assay, Luciferase, Enzyme-linked Immunosorbent Assay, Concentration Assay, Cell Culture

Journal: International Journal of Oncology

Article Title: Effects of non-steroidal anti-inflammatory drug-activated gene-1 on Ganoderma lucidum polysaccharides-induced apoptosis of human prostate cancer PC-3 cells

doi: 10.3892/ijo.2018.4578

Figure Lengend Snippet: GLP-induced apoptosis of PC-3 cells is mediated through NAG-1 induction. (A) NAG-1 siRNA successfully knocked down NAG-1 expression in PC-3 cells, as determined by western blotting. (B) NAG-1 siRNA inhibited GLP-induced NAG-1 expression, as determined by western blotting. (C) NAG-1 siRNA inhibited GLP-induced apoptosis, as determined by flow cytometry. Percentage of early and late apoptotic of PC-3 cells induced by GLP is presented in the lower panel. (D) NAG-1 siRNA inhibited GLP-induced PARP cleavage and the suppression of pro-caspase-3, -6 and -9 protein expression. β-actin was used as an internal control. Data are presented as the means ± standard error. * P<0.05, ** P<0.01 compared with the control group (one-way analysis of variance with Dunnett's correction). ## P<0.01, compared with GLP-treated cells. GLP, Ganoderma lucidum polysaccharides; NAG-1, non-steroidal anti-inflammatory drug-activated gene-1; PARP, poly(ADP-ribose) polymerase; PI, propidium iodide; siRNA, small interfering RNA.

Article Snippet: The Quantikine Human GDF15/NAG-1 ELISA kit (cat. no. DY957) was purchased from R&D Systems, Inc. (Minneapolis, MN, USA).

Techniques: Expressing, Western Blot, Flow Cytometry, Small Interfering RNA

Journal: International Journal of Oncology

Article Title: Effects of non-steroidal anti-inflammatory drug-activated gene-1 on Ganoderma lucidum polysaccharides-induced apoptosis of human prostate cancer PC-3 cells

doi: 10.3892/ijo.2018.4578

Figure Lengend Snippet: Working model of the molecular mechanisms by which GLP exerts its anticancer activity in prostate cancer PC-3 cells. GLP-induced apoptosis is mediated by NAG-1 induction, which may serve a pivotal role in GLP-induced cell death in prostate cancer cells.

Article Snippet: The Quantikine Human GDF15/NAG-1 ELISA kit (cat. no. DY957) was purchased from R&D Systems, Inc. (Minneapolis, MN, USA).

Techniques: Activity Assay

Journal: Cancers

Article Title: Visfatin Mediates Malignant Behaviors through Adipose-Derived Stem Cells Intermediary in Breast Cancer

doi: 10.3390/cancers12010029

Figure Lengend Snippet: GDF15 play a crucial role in the regulations of breast cancer cells activities by visfatin-treated ADSCs (vADSCs). ( A ) After the three-day co-culture of MDA-MB-231 cells and vADSCs (V50 and V100 group) or untreated ADSCs (Ctrl group), the CM was collected and analyzed by using a cytokine array kit. ( B ) The expression of GDF15 in the co-cultured CM was validated by ELISA represented in a histogram. The ADSCs and MDA-MB-231 cells collected from the co-culture system were extracted for cell lysate to analyze the GDF15 expression by western blotting. ( C ) The migration and invasion of MDA-MB-231 treated with GDF15 at various concentrations for 48 h were evaluated by using a transwell system. ( D ) The indirect co-culture was performed in the presence or absence of the GDF15 neutralizing antibody of for three days. After that, the migration and invasion of the MDA-MB-231 cells collected from the co-culture were evaluated in a transwell system. ( E ) The expression of phosphor-AKT (pAKT) of MDA-MB-231 treated with GDF15 (50 ng/mL) at different time point was detected by western blotting. ( F ) The pAKT was detected in the MDA-MB-231 cells from the co-culture by western blotting. ( G ) After the three-day co-culture in the presence or absence of the wortmannin (400 nM), the MDA-MB-231 cells were collected from the co-culture for performing the migration assay. All experiments were performed in triplicate.

Article Snippet: For performing cell invasion or migration assay, the MDA-MB-231 cells collected from the co-culture system or treated with human recombinant GDF15 (10936-H01H, Sino Biological, Wayne, PA, USA) were suspended in serum-free DMEM and transferred to the 24-well transwell inserts coated with or without Matrigel, respectively (8 μm pores, Corning).

Techniques: Co-Culture Assay, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot, Migration

Journal: Cancers

Article Title: Visfatin Mediates Malignant Behaviors through Adipose-Derived Stem Cells Intermediary in Breast Cancer

doi: 10.3390/cancers12010029

Figure Lengend Snippet: Visfatin-primed ADSCs promoted the tube formation of HUVEC. ( A ) HUVEC cells were co-cultured with visfatin-treated ADSCs or untreated ADSCs, noted as V50 and V100 or Ctrl, respectively, for three days. The HUVEC cells were collected from the co-culture and seeded in a matrix gel-coated 96-well plate. The tube formation of HUVEC was observed using a microscope. The length of branches was determined by using the ImageJ software. ( B ) After the three-day co-culture in the presence or absence of GDF15 neutralizing antibody (GDF15 Nab, 5 μg/mL), the HUVEC cells were collected from the co-culture for performing the tube formation assay. The experiments were performed in triplicate.

Article Snippet: For performing cell invasion or migration assay, the MDA-MB-231 cells collected from the co-culture system or treated with human recombinant GDF15 (10936-H01H, Sino Biological, Wayne, PA, USA) were suspended in serum-free DMEM and transferred to the 24-well transwell inserts coated with or without Matrigel, respectively (8 μm pores, Corning).

Techniques: Cell Culture, Co-Culture Assay, Microscopy, Software, Tube Formation Assay

Journal: Cancers

Article Title: Visfatin Mediates Malignant Behaviors through Adipose-Derived Stem Cells Intermediary in Breast Cancer

doi: 10.3390/cancers12010029

Figure Lengend Snippet: Visfatin-pretreated ADSCs (vADSCs) enhanced the tumor growth and metastasis in human breast cancer xenograft mouse model. ( A ) The nude mice were injected with mixture of MDA-MB-231 and untreated ADSCs (uADSCs) or vADSCs, noted as Ctrl or V50, respectively, to the mammary fat pads. The tumor volumes were measured every week after injection (Ctrl, n = 5; V50, n = 6). ( B ) After sacrificing the mice, the weight of the resected tumor was measured. ( C ) The expressions of GDF15, β-catenin, CD31, and pAKT in the tumor sections were detected by immunohistochemistry. The IHC score was calculated by multiplying the percentage of positive cells by the intensity and present as histogram. ( D ) The luciferase-expressing MDA-MB-231 were collected and injected into the tail vein of NOD/SCID mice after co-culturing with uADSCs or vADSCs, noted as Ctrl or V50, respectively (Ctrl, n = 8; V50, n = 8). The IVIS radiance signals of the mice were assessed at week 4. The representative images of high and low signal were shown. The statistical differences were calculated by t-test, *, p -value < 0.05; **, p -value < 0.01.

Article Snippet: For performing cell invasion or migration assay, the MDA-MB-231 cells collected from the co-culture system or treated with human recombinant GDF15 (10936-H01H, Sino Biological, Wayne, PA, USA) were suspended in serum-free DMEM and transferred to the 24-well transwell inserts coated with or without Matrigel, respectively (8 μm pores, Corning).

Techniques: Injection, Immunohistochemistry, Luciferase, Expressing

Journal: Cancers

Article Title: Visfatin Mediates Malignant Behaviors through Adipose-Derived Stem Cells Intermediary in Breast Cancer

doi: 10.3390/cancers12010029

Figure Lengend Snippet: The expressions of visfatin, GDF15, and pAKT in the specimens from breast cancer patients. ( A ) The expressions of visfatin, GDF15, and pAKT in breast cancer tissue microarray (n = 96) were detected by immunohistochemistry. The representative images of high expression levels (No. 1) and low expression levels (No. 2) were shown. The IHC score was calculated by multiplying the percentage of positive cells by the intensity, which was identified using HistoQuest Analysis Software. ( B ) The correlations between visfatin, GDF15, and pAKT according to the IHC score were calculated by using the online Pearson correlation coefficient calculator. ( C ) The correlation of serum levels of GDF15 and visfatin of breast cancer patients (n = 120) determined by ELISA was also calculated by using the online Pearson correlation coefficient calculator.

Article Snippet: For performing cell invasion or migration assay, the MDA-MB-231 cells collected from the co-culture system or treated with human recombinant GDF15 (10936-H01H, Sino Biological, Wayne, PA, USA) were suspended in serum-free DMEM and transferred to the 24-well transwell inserts coated with or without Matrigel, respectively (8 μm pores, Corning).

Techniques: Microarray, Immunohistochemistry, Expressing, Software, Enzyme-linked Immunosorbent Assay

Journal: Cancers

Article Title: Visfatin Mediates Malignant Behaviors through Adipose-Derived Stem Cells Intermediary in Breast Cancer

doi: 10.3390/cancers12010029

Figure Lengend Snippet: Visfatin mediates its effects both directly via cAbl/STAT3 and indirectly mediated by ADSCs via GDF15/AKT on promoting malignant behavior in breast cancer. Previously, we discovered visfatin mainly produced by adipocytes promoted breast cancer cells directly through activation of c-Abl and STAT3, which was blocked by Imatinib and Stattic inhibitor, respectively (black arrow). In this study, we showed that visfatin can act via an indirect pathway by priming ADSCs, which may be recruited from the adipose tissue to tumor site or generated from autologous fat transfer, to produce GDF15 that stimulated AKT activation in breast cancer cells to promote malignant behaviors (white arrow). The effect can be blocked by the treatment of GDF15 neutralizing Ab or Wortmannin inhibitor.

Article Snippet: For performing cell invasion or migration assay, the MDA-MB-231 cells collected from the co-culture system or treated with human recombinant GDF15 (10936-H01H, Sino Biological, Wayne, PA, USA) were suspended in serum-free DMEM and transferred to the 24-well transwell inserts coated with or without Matrigel, respectively (8 μm pores, Corning).

Techniques: Produced, Activation Assay, Generated