human gastric mucosal epithelial cells  (Procell Inc)

Journal: World Journal of Gastrointestinal Oncology

Article Title: Weiwei Decoction alleviates gastric intestinal metaplasia through the olfactomedin 4/nucleotide-binding oligomerization domain 1/caudal-type homeobox gene 2 signaling pathway

doi: 10.4251/wjgo.v16.i7.3211

Figure Lengend Snippet: Olfactomedin 4 exhibited direct binding and subsequent down-regulation of nucleotide-binding oligomerization domain 1, thereby sustaining the activation of caudal-type homeobox gene 2 and promoting the progression of intestinal metaplasia. A: Relative mRNA expressions of caudal-type homeobox gene 2 (CDX2), MUCIN 2 (MUC2), and villin 1 (VIL1) on stimulation of chenodeoxycholic acid (CDCA) in GES-1 cells ( n = 3); B: Western blot (WB) detection of MUC2, VIL1, and CDX2 in GES-1 cells treated with CDCA (100 μM) for 24 h; C: Timeline of the cell experiments design for pathway regulatory verification; D: Short hairpin RNAs (shRNAs) targeting olfactomedin 4 (OLFM4) were transfected to GES-1 cells. Subsequently, the cells were treated with CDCA (100 μM) for 24 h. WB detection of OLFM4, nucleotide-binding oligomerization domain 1 (NOD1), CDX2, and MUC2; E: AGS cells were transfected with OLFM4 shRNAs. WB detection of OLFM4, NOD1, CDX2, MUC2, and VIL1; F: OLFM4-pcDNAs were transfected to GES-1 cells. WB detection of OLFM4, NOD1, CDX2, and VIL1; G: Co-immunoprecipitation detection of the interaction between OLFM4 and NOD1, and the interaction between OLFM4 and CDX2 in GES-1 cells; H: OLFM4 and NOD1 shRNAs were co-transfected to GES-1 cells. Subsequently, the cells were treated with CDCA (100 μM) for 24 h. WB detection of OLFM4, NOD1, CDX2, and MUC2. a P < 0.05, b P < 0.01, c P < 0.001 vs control group. OLFM4: Olfactomedin 4; NOD1: Nucleotide-binding oligomerization domain 1; CDX2: Caudal-type homeobox gene 2; MUC2: MUCIN 2; VIL1: Villin 1; CDCA: Chenodeoxycholic acid.

Article Snippet: Human gastric mucosal epithelial cell-line GES-1 (Procell, Wuhan, China) and human gastric adenocarcinoma cell-line AGS (ATCC, Manassas, VA) were cultured in RPMI-1640 medium (Gibco, United States) supplemented with 10% (V/V) fetal bovine serum (Gibco, United States) in a humidified incubator at 37 °C and 50 mL/L CO 2 .

Techniques: Binding Assay, Activation Assay, Western Blot, Transfection, Immunoprecipitation, Control

Journal: World Journal of Gastrointestinal Oncology

Article Title: Weiwei Decoction alleviates gastric intestinal metaplasia through the olfactomedin 4/nucleotide-binding oligomerization domain 1/caudal-type homeobox gene 2 signaling pathway

doi: 10.4251/wjgo.v16.i7.3211

Figure Lengend Snippet: Weiwei Decoction-medicated serum attenuates olfactomedin 4 level and simultaneously strengthens olfactomedin 4 and nucleotide-binding oligomerization domain 1 binding, thereby enhancing nucleotide-binding oligomerization domain 1 expression and consequently mitigating caudal-type homeobox gene 2 in gastric cells. A: Timeline of the cell experiments design for Weiwei Decoction (WWD)-medicated serum treatment; B: GES-1 cells were treated with chenodeoxycholic acid (CDCA) for 24 h and then disposed to 10% (V/V) of the corresponding WWD-medicated serum for 48 h. WB detection of olfactomedin 4 (OLFM4), nucleotide-binding oligomerization domain 1 (NOD1), caudal-type homeobox gene 2 (CDX2), and MUCIN 2 (MUC2); C: Relative protein expression of OLFM4, NOD1, CDX2, and MUC2 in GES-1 cells ( n = 3); D: AGS cells were disposed to 10% (V/V) of the corresponding WWD-medicated serum for 48 h. WB detection of OLFM4, NOD1, CDX2, MUC2, and villin 1 (VIL1); E: Relative protein expression of OLFM4, NOD1, CDX2, MUC2, and VIL1 in AGS cells ( n = 3); F: GES-1 cells were treated with CDCA for 24 h and then disposed to 10% (V/V) of the WWD-H medicated serum for 48 h. Co-immunoprecipitation detection of the interaction between OLFM4 and NOD1, and the interaction between OLFM4 and CDX2 in GES-1 cells. a P < 0.05, b P < 0.01 vs control group; c P < 0.05, d P < 0.01 vs model group. OLFM4: Olfactomedin 4; NOD1: Nucleotide-binding oligomerization domain 1; CDX2: Caudal-type homeobox gene 2; WWD: Weiwei Decoction; MUC2: MUCIN 2; VIL1: Villin 1.

Article Snippet: Human gastric mucosal epithelial cell-line GES-1 (Procell, Wuhan, China) and human gastric adenocarcinoma cell-line AGS (ATCC, Manassas, VA) were cultured in RPMI-1640 medium (Gibco, United States) supplemented with 10% (V/V) fetal bovine serum (Gibco, United States) in a humidified incubator at 37 °C and 50 mL/L CO 2 .

Techniques: Binding Assay, Expressing, Immunoprecipitation, Control

Journal: World Journal of Gastrointestinal Oncology

Article Title: Weiwei Decoction alleviates gastric intestinal metaplasia through the olfactomedin 4/nucleotide-binding oligomerization domain 1/caudal-type homeobox gene 2 signaling pathway

doi: 10.4251/wjgo.v16.i7.3211

Figure Lengend Snippet: Weiwei Decoction inhibits cytokines and chemokines in gastric intestinal metaplasia. A: Enzyme-linked immunosorbent assay was utilized to detect interferon-gamma, interleukin (IL)-6, IL-17, macrophage chemoattractant protein-1, and macrophage inflammatory protein 1 alpha in intestinal metaplasia rats serum ( n = 6); B: Relative mRNA expression of tumor necrosis factor alpha, IL-6, and IL-8 in GES-1 cells ( n = 3). a P < 0.05, b P < 0.01, c P < 0.001 vs control group; d P < 0.05, e P < 0.01, f P < 0.001 vs model group; g P < 0.05, h P < 0.01, i P < 0.001 vs folic acid group. IL: Interleukin; MIP: Macrophage inflammatory protein; MCP: Macrophage chemoattractant protein; IFN: Interferon; WWD: Weiwei Decoction; FA: Folic acid; TNF: Tumor necrosis factor.

Article Snippet: Human gastric mucosal epithelial cell-line GES-1 (Procell, Wuhan, China) and human gastric adenocarcinoma cell-line AGS (ATCC, Manassas, VA) were cultured in RPMI-1640 medium (Gibco, United States) supplemented with 10% (V/V) fetal bovine serum (Gibco, United States) in a humidified incubator at 37 °C and 50 mL/L CO 2 .

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Control