human gc cell lines snu 16  (ATCC)

Journal: eLife

Article Title: Blocking SHP2 benefits FGFR2 inhibitor and overcomes its resistance in FGFR2 -amplified gastric cancer

doi: 10.7554/eLife.104060

Figure Lengend Snippet: Sensitivity of KATOIII and SNU-16 to ( A ) SHP099, ( B ) AZD4547, or combination therapy with different concentration gradients (n=4; two-way ANOVA). Effects of different treatments on cell apoptosis of ( C ) KATOIII and ( D ) SNU-16 after 48-hour drugs incubation (n=3; one-way ANOVA). ( E ) KATOIII and ( F ) SNU-16 were incubated with vehicle, SHP099 10 μM, AZD4547 3 nM or combination therapies for 1 hour or 48 hours, then cell lysates were immunoblotted for phospho-FGFR and total-FGFR2, phospho-SHP2 and total-SHP2, phospho-Erk and total-Erk, phospho-p38 and total-p38, phospho-AKT and total-AKT, and phospho-mTOR and total-mTOR. Data are shown as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. p-Values are determined by ordinary one-way ANOVA or two-way ANOVA. Figure 2—source data 1. PDF file containing original western blots for , indicating the relevant bands. Figure 2—source data 2. Original file for western blots displayed in .

Article Snippet: Human GC cell lines SNU-16 (ATCC CRL-5974), MKN45 (KANGBAI CBP60488), NUGC4 (KANGBAI CBP60493), HGC27 (KANGBAI CBP60480), and SNU601 (KANGBAI CBP60507) were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China).

Techniques: Concentration Assay, Incubation, Western Blot

Journal: eLife

Article Title: Blocking SHP2 benefits FGFR2 inhibitor and overcomes its resistance in FGFR2 -amplified gastric cancer

doi: 10.7554/eLife.104060

Figure Lengend Snippet: BALB/c nude mice were injected with 1×10 7 SNU-16 cancer cells and received different formulations by oral gavage every day upon tumor volumes reached 100–150 mm 3 . ( A ) Schematic of SHP099 and AZD4547 therapeutic schedule in FGFR2 -amplified SNU-16 subcutaneous xenograft model. ( B ) Representative images of tumors (n=5). ( C ) Tumor volumes (n=5; two-way ANOVA), ( D ) tumor weights (n=5; one-way ANOVA) and ( E ) body weights (n=5; two-way ANOVA) of different groups. ( F ) Tumors were harvested 6 hours after the last dose of drugs, and cell lysates from tumor tissues were immunoblotted for phospho-FGFR and total-FGFR2, phospho-SHP2 and total-SHP2, phospho-Erk and total-Erk, phospho-AKT and total-AKT, and phospho-mTOR and total-mTOR. Data are shown as mean ± SEM. ns, not significant, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. p-Values are determined by ordinary one-way ANOVA or two-way ANOVA. Figure 4—source data 1. PDF file containing original western blots for , indicating the relevant bands. Figure 4—source data 2. Original file for western blots displayed in .

Article Snippet: Human GC cell lines SNU-16 (ATCC CRL-5974), MKN45 (KANGBAI CBP60488), NUGC4 (KANGBAI CBP60493), HGC27 (KANGBAI CBP60480), and SNU601 (KANGBAI CBP60507) were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China).

Techniques: Injection, Amplification, Western Blot

Journal: eLife

Article Title: Blocking SHP2 benefits FGFR2 inhibitor and overcomes its resistance in FGFR2 -amplified gastric cancer

doi: 10.7554/eLife.104060

Figure Lengend Snippet: ( A ) Overview of FGFR2 alterations individually in GC patients from Nanjing Drum Tower hospital cohort. ( B ) PD-L1 mRNA expression levels were analyzed between FGFR2 -amplified group (n=13) and FGFR2 -unamplified group (n=250) from TCGA-STAD cohort (Welch’s t -test). ( C ) Proportions of MSI-H or MSS in FGFR2 -amplified group (n=12) and FGFR2 -unamplified group (n=237) from TCGA-STAD cohort (Fisher’s exact test). Human peripheral blood mononuclear cells (PBMCs) were incubated with different administrations in the presence of 0.25 μg/ml human anti-CD3 and 1 μg/ml human anti-CD28. Proportions of ( D ) IFN-γ/CD8+ cells were detected by flow cytometry assay after 24 hours of drugs incubation (n=3; one-way ANOVA). ( E ) Expression levels of IFN-γ in cellular supernatant were measured by Cytometric Bead Array (CBA) assay after 24 hours of drugs incubation (n=3; one-way ANOVA). ( F ) Cell viability of human PBMCs after 48 hours of drugs incubation (n=4; one-way ANOVA). ( G, H ) After 48 hours of advance drugs stimulation in human PBMCs, cytotoxicities of human PBMCs against SNU-16 at different E:T ratios of 10:1, 20:1, 40:1 were analyzed by flow cytometry assay after CFSE/PI staining (n=3; two-way ANOVA). Data are shown as mean ± SEM. ns, not significant, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. p-Values are determined by Welch’s t -test, Fisher’s exact test, ordinary one-way ANOVA or two-way ANOVA.

Article Snippet: Human GC cell lines SNU-16 (ATCC CRL-5974), MKN45 (KANGBAI CBP60488), NUGC4 (KANGBAI CBP60493), HGC27 (KANGBAI CBP60480), and SNU601 (KANGBAI CBP60507) were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China).

Techniques: Expressing, Amplification, Incubation, Flow Cytometry, Staining

Journal: Frontiers in Genetics

Article Title: Systematic analysis of anoikis-related genes identifies SRPX2-FAK/AKT-IL-6 axis in the progression and peritoneal metastasis of gastric cancer

doi: 10.3389/fgene.2025.1736097

Figure Lengend Snippet: SRPX2 in CAFs promotes malignant GC progression in vitro and in vivo . (A,B) Transwell migration assay and quantitative analysis of HGC and AGS cells transfected with siSRPX2#1 or siSRPX2#2. (C) Immunofluorescence staining showing subcellular localization and expression of α-SMA in isolated CAFs and NFs. (D) qRT-PCR analysis of SRPX2 expression levels in isolated CAFs and NFs. (E,F) Colony formation assay of HGC and AGS cells co-cultured with PBS, NFs, or CAFs. (G,H) Transwell migration assay of HGC and AGS cells co-cultured with PBS, NFs, or CAFs. (I) Wound healing assay of HGC and AGS cells co-cultured with PBS, NFs, or CAFs. (J) Flow cytometry analysis of apoptosis in AGS cells under anoikis conditions (low-attachment culture) after co-culture with PBS, NFs, or CAFs. (K–O) Colony formation, Transwell migration, and wound healing assays of HGC and AGS cells cultured with conditioned medium from CAFs transfected with siSRPX2#1 or siSRPX2#2. (P) Flow cytometry analysis of apoptosis rates in AGS cells cultured with conditioned medium from CAFs transfected with siSRPX2#1 or siSRPX2#2. (Q–T) In vivo xenograft tumor assay: (Q) representative images of tumors from mice co-injected with MKN45 cells and CAFs (n = 5) or MKN45 cells and SRPX2-knockdown CAFs (shSRPX2, n = 5); (R) tumor growth curves; (S) average tumor weight; (T) representative H&E staining and Ki-67 immunohistochemistry of xenograft tumor sections. All data are presented as mean ± SD of triplicate experiments. ns, P > 0.05; ***, P < 0.001.

Article Snippet: The GC cell lines AGS, HGC-27, and MKN45 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, United States) and maintained in the Gastrointestinal Oncology Laboratory at Sun Yat-sen Memorial Hospital, Sun Yat-sen University.

Techniques: In Vitro, In Vivo, Transwell Migration Assay, Transfection, Immunofluorescence, Staining, Expressing, Isolation, Quantitative RT-PCR, Colony Assay, Cell Culture, Wound Healing Assay, Flow Cytometry, Co-Culture Assay, Migration, Injection, Knockdown, Immunohistochemistry

Journal: Frontiers in Genetics

Article Title: Systematic analysis of anoikis-related genes identifies SRPX2-FAK/AKT-IL-6 axis in the progression and peritoneal metastasis of gastric cancer

doi: 10.3389/fgene.2025.1736097

Figure Lengend Snippet: SRPX2 in CAFs activates the FAK/AKT pathway and secretes IL-6 to induce anoikis resistance in GC. (A) Western blot analysis of SRPX2, p-FAK, FAK, p-AKT, and AKT expression in CAFs transfected with siSRPX2#1 or siSRPX2#2. (B) Expression levels of p-FAK and p-AKT in CAFs overexpressing SRPX2 and treated with the FAK pathway inhibitor PF-573228 (5 μM). (C) Expression of SRPX2, p-FAK, and p-AKT in CAFs overexpressing TFAP2A, followed by SRPX2 knockdown. (D,E) Relative IL-6 mRNA expression in CAFs after SRPX2 knockdown or overexpression, measured by qRT-PCR. (F,G) IL-6 secretion levels in conditioned medium from CAFs after SRPX2 knockdown or overexpression, as determined by ELISA. (H) Apoptosis analysis by Annexin V/PI staining in AGS cells cultured under anoikis conditions (low-attachment culture) after treatment with conditioned medium from SRPX2-overexpressing CAFs pretreated with tocilizumab (10 μg/mL) for 24 h. (I,J) Tocilizumab treatment significantly suppressed subcutaneous tumor growth in BALB/c nude mice injected with MKN45 cells and SRPX2-overexpressing CAFs. (K) In vivo monitoring of GC cell dissemination in the abdominal cavity using IVIS imaging; tocilizumab treatment markedly inhibited peritoneal metastasis. (L) Representative images of peritoneal metastases in mice at the end of the study. All data are presented as mean ± SD of triplicate experiments. ***, P < 0.001.

Article Snippet: The GC cell lines AGS, HGC-27, and MKN45 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, United States) and maintained in the Gastrointestinal Oncology Laboratory at Sun Yat-sen Memorial Hospital, Sun Yat-sen University.

Techniques: Western Blot, Expressing, Transfection, Knockdown, Over Expression, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Staining, Cell Culture, Injection, In Vivo, Imaging