gastric cancer cell line ags mgc803  (ATCC)

Journal: Discover Oncology

Article Title: Rutin suppresses the malignant biological behavior of gastric cancer cells through the Wnt/β-catenin pathway

doi: 10.1007/s12672-024-01281-w

Figure Lengend Snippet: Effects of Rutin on cell proliferation and apoptosis in AGS and MGC803 cells. A , B Detection of cellular viability with MTT assays; C , D Flow cytometry detected the cell apoptosis; E – H The protein level of cleaved-Caspase3 were determined with western blotting. * p < 0.05; ** p < 0.01

Article Snippet: Gastric cancer cell line AGS\MGC803 and the normal gastric epithelial cell line GES-1 were purchased from American Type Culture Collection (ATCC).

Techniques: Flow Cytometry, Western Blot

Journal: Discover Oncology

Article Title: Rutin suppresses the malignant biological behavior of gastric cancer cells through the Wnt/β-catenin pathway

doi: 10.1007/s12672-024-01281-w

Figure Lengend Snippet: Effects of Rutin on cell migration in AGS and MGC803 cells. A , B Scratch assays were used to evaluate migration AGS and MGC803 cells. * p < 0.05; ** p < 0.01

Article Snippet: Gastric cancer cell line AGS\MGC803 and the normal gastric epithelial cell line GES-1 were purchased from American Type Culture Collection (ATCC).

Techniques: Migration

Journal: Discover Oncology

Article Title: Rutin suppresses the malignant biological behavior of gastric cancer cells through the Wnt/β-catenin pathway

doi: 10.1007/s12672-024-01281-w

Figure Lengend Snippet: Effects of Rutin on cell invasion in AGS and MGC803 cells. A , B Transwell assays were used to evaluate invasion of AGS and MGC803 cells. ** p < 0.01

Article Snippet: Gastric cancer cell line AGS\MGC803 and the normal gastric epithelial cell line GES-1 were purchased from American Type Culture Collection (ATCC).

Techniques:

Journal: Discover Oncology

Article Title: Rutin suppresses the malignant biological behavior of gastric cancer cells through the Wnt/β-catenin pathway

doi: 10.1007/s12672-024-01281-w

Figure Lengend Snippet: Effects of Rutin on EMT in AGS and MGC803 cells. A – C Western blotting and RT-qPCR detected protein and mRNA levels of E-cadherin and N-cadherin in AGS cells. D–F Western blotting and RT-qPCR detected protein and mRNA levels of E-cadherin and N-cadherin in MGC803 cells. * p < 0.05; ** p < 0.01

Article Snippet: Gastric cancer cell line AGS\MGC803 and the normal gastric epithelial cell line GES-1 were purchased from American Type Culture Collection (ATCC).

Techniques: Western Blot, Quantitative RT-PCR

Journal: Discover Oncology

Article Title: Rutin suppresses the malignant biological behavior of gastric cancer cells through the Wnt/β-catenin pathway

doi: 10.1007/s12672-024-01281-w

Figure Lengend Snippet: Rutin inhibits Wnt/β-catenin signaling in AGS and MGC803 cells. A – C . Western blotting and RT-qPCR detected protein and mRNA levels of Wnt3a and β-catenin in AGS cells. D – F Western blotting and RT-qPCR detected protein and mRNA levels of Wnt3a and β-catenin in MGC803 cells. * p < 0.05; ** p < 0.01

Article Snippet: Gastric cancer cell line AGS\MGC803 and the normal gastric epithelial cell line GES-1 were purchased from American Type Culture Collection (ATCC).

Techniques: Western Blot, Quantitative RT-PCR

Journal: Translational Oncology

Article Title: Collagen extracellular matrix promotes gastric cancer immune evasion by activating IL4I1-AHR signaling

doi: 10.1016/j.tranon.2024.102113

Figure Lengend Snippet: Collagen ECM-cultured tumor cells promote CAR-T cell exhaustion A, Graphical representation of short-term specific cytotoxicity experiment in vitro . Briefly, 5 × 104 Her2+ MGC803 or HGC27 which was pre-cultured on gels of different dose collagen co-cultured with CAR T cells at an E:T ratio of 1:1 for two days. At the end of the first round of co-culture, the viable CAR-T cells were counted and the CAR-T-mediated specific cytotoxicity was detected by Annexin V-FITC/PI Apoptosis Detection Kit. B, Cytotoxic activity of HER2-specific chimeric antigen receptor-T cells against MGC803 or HGC27. C-D, the expression of multiple inhibitory receptors (IRs) (C) and cytokines (D) of HER2-specific chimeric antigen receptor-T cells against MGC803 or HGC27. E-F, Graphical representation of long-term specific cytotoxicity experiment in vitro . Briefly, 5 × 104 Her2+ MGC803 or HGC27 which pre-cultured on dish or collagen gel co-cultured with 2D-CAR or 3D-CAR T cells at an E:T ratio of 1:1 for two days. At the end of the first round of co-culture, the viable CAR-T cells were counted and the CAR-T-mediated specific cytotoxicity was detected by Annexin V-FITC/PI Apoptosis Detection Kit, and a second round of co-culture using the first round of CAR+ T cells with the same number of tumor cells at the same E:T ratio (1:1). a second round of co-culture assays were performed (F). n = 3 biological independent experiments (A to F). **P < 0.01, ***P < 0.001 or NS (no significant difference, by two-tailed Student's t test (C, D and F) or one-way ANOVA and Bonferroni's test (B). The data represent means ± SD.

Article Snippet: Human gastric cancer cell line MGC803, HGC27 and HEK293T (X100478) were purchased from the China Center for Type Culture Collection (Beijing, China).

Techniques: Cell Culture, In Vitro, Co-Culture Assay, Activity Assay, Expressing, Two Tailed Test

Journal: Translational Oncology

Article Title: Collagen extracellular matrix promotes gastric cancer immune evasion by activating IL4I1-AHR signaling

doi: 10.1016/j.tranon.2024.102113

Figure Lengend Snippet: IL4I1 is upregulated in Collagen ECM-cultured tumor cells A, Immunofluorescence analysis of IL4I1 expression in MGC803 or HGC27 derived from the 2D and 3D groups. Scale bar, 20 μm. B-C, The cytotoxic activity of HER2-specific chimeric antigen receptor-T cells against MGC803 or HGC27 which were transfected with sgNC, sgIL4I1-1 and sgIL4I1-2. C, NSG mice were transplanted with PDX. At 30 days post engraftment, NSG mice were adoptively transferred with 1 × 106 human HER2-CAR or Mock T cells, then at 30 days post engraftment, mice were taken out in each group and killed for assessment of tumor weight(C). D-E, the expression of multiple IRs(D) and cytokines(E) of HER2-specific chimeric antigen receptor-T cells in PDX tissue. n = 3 biological independent experiments (B) and n = 6 biological independent experiments (C to E). **P < 0.01, ***P < 0.001 or NS (no significant difference, by one-way ANOVA and Bonferroni's test (B to E). The data represent means ± SD.

Article Snippet: Human gastric cancer cell line MGC803, HGC27 and HEK293T (X100478) were purchased from the China Center for Type Culture Collection (Beijing, China).

Techniques: Cell Culture, Immunofluorescence, Expressing, Derivative Assay, Activity Assay, Transfection

Journal: Translational Oncology

Article Title: Collagen extracellular matrix promotes gastric cancer immune evasion by activating IL4I1-AHR signaling

doi: 10.1016/j.tranon.2024.102113

Figure Lengend Snippet: 3D collagen induces IL4I1 expression through integrin αvβ1, thereby promoting tumor immune evasion A, qPCR analysis of Integrins expression in MGC803 or HGC27 derived from the 2D and 3D groups. B, Immunofluorescence analysis of integrin αvβ1 expression in MGC803 or HGC27 derived from the 2D and 3D groups. Scale bar, 20 μm. C, Immunofluorescence analysis of IL4I1 expression in MGC803 or HGC27 treated with or without integrin αvβ1 inhibitor. Scale bar, 20 μm. D, Her2+ MGC803 or HGC27 which was treated with or without integrin αvβ1 inhibitor co-cultured with CAR-T cells at an E:T ratio of 1:1 for two days. CAR-T-mediated specific cytotoxicity was detected by Annexin V-FITC/PI Apoptosis Detection Kit. E-F, the expression of multiple IRs (E) and cytokines (F) of HER2-specific chimeric antigen receptor-T cells against MGC803 or HGC27 which was treated with or without integrin αvβ1 inhibitor. G-I, NSG mice were transplanted with PDX. At 30 days post-engraftment, NSG mice were adoptively transferred with 1 × 106 human HER2-CAR or Mock T cells, combined with either PBS or integrin αvβ1 inhibitor. The expression of IRs (G) and cytokines (H) of HER2-specific chimeric antigen receptor-T cells in PDX tissue. then at 60 days post-engraftment, mice were taken out in each group and killed for assessment of tumor weight (I). n = 3 biological independent experiments (A to F) and n = 6 biological independent experiments (G to H). **P < 0.01, ***P < 0.001 or NS (no significant difference, by Two-tailed Student's t-test. The data represent means ± SD.

Article Snippet: Human gastric cancer cell line MGC803, HGC27 and HEK293T (X100478) were purchased from the China Center for Type Culture Collection (Beijing, China).

Techniques: Expressing, Derivative Assay, Immunofluorescence, Cell Culture, Two Tailed Test

Journal: Translational Oncology

Article Title: Collagen extracellular matrix promotes gastric cancer immune evasion by activating IL4I1-AHR signaling

doi: 10.1016/j.tranon.2024.102113

Figure Lengend Snippet: 3D collagen gel promotes tumor immune evasion through the integrin αvβ1/AKT/SP1/IL4I1 signaling pathway. A, Immunofluorescence analysis of p-AKT expression in MGC803 or HGC27 treated with or without integrin αvβ1 inhibitor. Scale bar, 20 μm. B, Immunofluorescence analysis of p-AKT expression in MGC803 or HGC27 treated with or without AKT inhibitor. Scale bar, 20 μm. C, Her2+ MGC803 or HGC27 which was treated with or without AKT inhibitor co-cultured with CAR-T cells at an E:T ratio of 1:1 for two days. CAR-T-mediated specific cytotoxicity was detected by Annexin V-FITC/PI Apoptosis Detection Kit. D, Immunofluorescence analysis of SP1 expression in MGC803 or HGC27 treated with or without AKT inhibitor and integrin αvβ1 inhibitor. Scale bar, 20 μm. E, Immunofluorescence analysis of IL4I1 expression in MGC803 or HGC27 transfected with or without sgSP1. Scale bar, 20 μm. F, MGC803 or HGC27 derived from the 2D and 3D groups. ChIP–qPCR analysis was performed with an antibody to SP1 and IL4I1-promotor-specific primers. G, MGC803 cells were cotransfected with a IL4I1 promoter-luciferase reporter PGL3 and Flag–SP1 plasmid for 24 h. Cells were then cultured with 2D or 3D gel for another 48 h, followed by analysis of luciferase activity. H-J, NSG mice were transplanted with PDX transfected with or without sgSP1. At 30 days post engraftment, NSG mice were adoptively transferred with 1 × 106 human HER2-CAR or Mock T cells, then at 30 days post engraftment, mice were taken out in each group and killed for the expression of IRs (H), cytokines (I) and assessment of tumor weight (J).n = 3 biological independent experiments (A to G) and n = 6 biological independent experiments (H to J). **P < 0.01, ***P < 0.001 or NS (no significant difference, by one-way ANOVA and Bonferroni's test (F to J). Two-tailed Student's t-test (C). The data represent means ± SD.

Article Snippet: Human gastric cancer cell line MGC803, HGC27 and HEK293T (X100478) were purchased from the China Center for Type Culture Collection (Beijing, China).

Techniques: Immunofluorescence, Expressing, Cell Culture, Transfection, Derivative Assay, Luciferase, Plasmid Preparation, Activity Assay, Two Tailed Test

Journal: Translational Oncology

Article Title: Collagen extracellular matrix promotes gastric cancer immune evasion by activating IL4I1-AHR signaling

doi: 10.1016/j.tranon.2024.102113

Figure Lengend Snippet: Collagen ECM-cultured tumor cells derived KynA promotes CAR-T exhaustion A, KynA levels measured by LC-MS in lysates of MGC803 or HGC27 derived from the 2D and 3D groups. B, qPCR analysis of tryptophan metabolism related enzymes expression in MGC803 or HGC27 derived from the 2D and 3D groups. C, KynA levels measured by LC-MS in lysates of MGC803 or HGC27 which were transfected with sgNC, sgIL4I1-1 and sgIL4I1-2. D-E, the expression of IRs (D) and cytokines (E) of HER2-specific chimeric antigen receptor-T cells which was treated with KynA. n = 3 biological independent experiments (A to E). **P < 0.01, ***P < 0.001 or NS (no significant difference, by one-way ANOVA and Bonferroni's test (C). Two-tailed Student's t-test (A,B,D,E). The data represent means ± SD.

Article Snippet: Human gastric cancer cell line MGC803, HGC27 and HEK293T (X100478) were purchased from the China Center for Type Culture Collection (Beijing, China).

Techniques: Cell Culture, Derivative Assay, Liquid Chromatography with Mass Spectroscopy, Expressing, Transfection, Two Tailed Test