ags human gc cell line  (ATCC)

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