Journal: World Journal of Surgical Oncology

Article Title: Analysis of the proliferative role and prognostic value of GPR173 in gastric cancer

doi: 10.1186/s12957-026-04274-x

Figure Lengend Snippet: Impact of GPR173 on gastric cancer cell proliferation in vitro and in vivo. A Relative mRNA expression of GPR173 in 10 pairs of gastric cancer tissues (Tumor) and adjacent Non-tumor tissues (Non-tumor) was measured by RT-qPCR. B Protein expression level of GPR173 in the human Non-tumor gastric mucosal epithelial cell line (GES-1) and various gastric cancer cell lines was detected by Western blot. C Western blot analysis validating the knockdown efficiency of GPR173 in MKN-45 cells and overexpression efficiency in AGS cells at the protein level. GAPDH served as the loading control. D The relative mRNA expression of GPR173 in MKN-45 cells transfected with different siRNAs (si-1, si-2, si-3) compared to negative control (NC) was determined by RT-qPCR. E The relative mRNA expression of GPR173 in AGS cells transfected with GPR173 overexpression plasmid (OE) compared to empty vector (Vector) was confirmed by RT-qPCR. F , H A colony formation assay and its quantification showed that knockdown of GPR173 inhibited the colony-forming ability of MKN-45 cells. G , H A colony formation assay and its quantification indicated that overexpression of GPR173 (OE) enhanced the colony-forming ability of AGS cells. I , K An EdU incorporation assay and its quantification revealed that knockdown of GPR173 suppressed the proliferation of MKN-45 cells. J , K An EdU incorporation assay and its quantification demonstrated that overexpression of GPR173 promoted the proliferation of AGS cells. L A CCK-8 assay showed that GPR173 knockdown inhibited the viability of MKN-45 cells. M A CCK-8 assay indicated that GPR173 overexpression enhanced the viability of AGS cells. N Representative images of subcutaneous xenograft tumors in nude mice injected with GPR173-overexpressing AGS cells (OE-GPR173) or control cells (Vector). O Quantitative comparison of tumor weights from each group at the experimental endpoint. P Tumor growth curves showing changes in tumor volume over time for each group. Q Representative images of IHC staining for GPR173 and the proliferation marker Ki-67 in subcutaneous xenograft tumors. Data are presented as the mean ± standard deviation (SD). Statistical significance was determined using Student’s t-test for comparisons between two groups A , D - O and one-way ANOVA for multiple comparisons. Tumor growth curves P were analyzed using two-way ANOVA

Article Snippet: The human Non-tumor gastric epithelial cell line GES-1 and human GC cell lines (MKN-45, HGC-27, SNU-216, MKN-73, AGS) were obtained from Genechem (Shanghai, China) or the Chinese Academy of Sciences Cell Bank (Shanghai, China).

Techniques: In Vitro, In Vivo, Expressing, Quantitative RT-PCR, Western Blot, Knockdown, Over Expression, Control, Transfection, Negative Control, Plasmid Preparation, Colony Assay, CCK-8 Assay, Injection, Comparison, Immunohistochemistry, Marker, Standard Deviation

Journal: MedComm

Article Title: Nitrate Enhances Gastric Mucosa Defense and Repair Process in Ethanol‐Induced Gastric Ulcer Rats via the Notch–Tff2 Pathway

doi: 10.1002/mco2.70628

Figure Lengend Snippet: Nitrate promotes migration by TFF2 upregulation in vitro. (A) Images of the scratch healing process of GES‐1 cells in Ibidi culture inserts. Scale bar = 500 µm. (B and C) Quantitative analysis of the migration rate at 24 h and 48 h in (A). (D and E) Representative immunoblotting band of the TFF2 protein and analysis of band gray values. (F) IF staining of pMLC (pink) and DAPI (blue). Scale bar = 40 µm. (G) IF analysis of pMLC with MFI. (H) Images of the scratch healing process of GES‐1 cells transfected with si‐ TFF2 /si‐negative control in Ibidi culture‐inserts. Scale bar = 500 µm. (I and J) Quantitative analysis of the migration rate at 24 h and 48 h. (K) Images of the scratch healing process of GES‐1 cells transfected with si‐ TFF2 /si‐negative control in Ibidi culture‐inserts containing ethanol. Scale bar = 500 µm. (L and M) Quantitative analysis of the migration rate at 24 h and 48 h. Migration rate is quantified by the percentage of closed area to the initial scratch area. Quantitative data are expressed as the mean ± SD. * p < 0.05, ** p < 0.01, and ns denotes no significance. TFF2, trefoil factor 2; GES‐1, human gastric epithelial; EtOH, ethanol; Nit, nitrate; Ctrl, control; MLC, myosin light chain 2; pMLC, phosphorylated myosin light chain 2; DAPI, 2‐(4‐amidinophenyl)‐6‐indolecarbamidine dihydrochloride; NC, negative control; SD, standard deviation.

Article Snippet: The in situ PLA was performed on fixed GES‐1 cells following the manufacturer's protocol (DUO82049, DUO92008; Merck).

Techniques: Migration, In Vitro, Western Blot, Staining, Transfection, Negative Control, Control, Standard Deviation

Journal: MedComm

Article Title: Nitrate Enhances Gastric Mucosa Defense and Repair Process in Ethanol‐Induced Gastric Ulcer Rats via the Notch–Tff2 Pathway

doi: 10.1002/mco2.70628

Figure Lengend Snippet: Nitrate functions by Notch pathway inhibition positively transcripting TFF2 in vitro. (A) Diagram of the wild‐type and mutant sequences for the three predicted RBPJ binding sites within the 251 bp TFF2 promoter probes. (B) Representative electrophoretic mobility shift assay (EMSA) autoradiograph. Hot probe is the biotin‐labeled wild‐type oligonucleotides of the truncated TFF2 promoter containing the binding motif; Mutant probe is the labeled oligonucleotides sequence with nucleotides mutated. The cold probe is nonlabeled competitive wild‐type probes (100 and 50 that of the concentrations). The shifted bands are indicated by arrows, which suggested the formation of DNA–protein complexes (lane 2, 3, 7). The super shifted bands indicated the formation of DNA–protein–antibody complexes (lane 3, 7). “+” and “−” represent presence and absence, respectively. (C) Relative TFF2 promoter (Full, Mut1, Mut2, and Mut3) luciferase activity was detected by DLR assays in RBPJ overexpressed and normal‐expressed GES‐1 cells. (D) A schematic diagram showing the location of RBPJ putative binding regions on the TFF2 promoter. (E) RT‐qPCR analysis of TFF2 binding site expression of GES‐1 cells. Target site expression in RBPJ‐treated groups was normalized to IgG negative control groups and expressed as fold change relative to the IgG groups. (F) IF staining of NICD (pink) and DAPI (blue). Scale bar = 50 µm. (G) IF analysis of MFI of nuclear NICD in positive cells. (H) Representative immunoblotting band of Notch signaling pathway in GES‐1 cells. (I–K) Analyses of immunoblotting band gray value of (H). (L) RT‐qPCR analysis of TFF2 mRNA expression of DMSO/DAPT treated GES‐1 cells. Target gene expression was normalized to GAPDH mRNA and expressed as fold change relative to the DMSO vehicle group. (M) RT‐qPCR analysis of TFF2 mRNA expression in RBPJ overexpressed and NICD deprived GES‐1 cells. The target gene expression was normalized to GAPDH mRNA and expressed as fold change relative to the vector + DMSO group. (N) RT‐qPCR analysis of TFF2 mRNA expression in RBPJ overexpressed and NICD‐RBPJ overexpressed GES‐1 cells. The target gene expression was normalized to GAPDH mRNA and expressed as fold change relative to the vector1 + vector2 group. (O) Representative image of PLA of NICD–RBPJ proximity. Each red dot represents a positive signal of NICD–RBPJ interaction and nuclei were counterstained with DAPI (blue). Scale bar = 20 µm. Quantitative data are expressed as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, and ns denotes no significance. Nit, nitrate; EtOH, ethanol; GES‐1, human gastric epithelial; NICD, intracellular structural domain; RBPJ, recombination signal binding protein for immunoglobulin kappa J region; EMSA, electrophoretic mobility shift assay; TFF2, trefoil factor 2; DAPI, 2‐(4‐amidinophenyl)‐6‐indolecarbamidine dihydrochloride; Yhhu‐3792, N2‐(4‐isopropylphenyl)‐5‐(3‐methoxyphenoxy) quinazoline‐2,4‐diamine; Luc, luciferase; SD, standard deviation; DLR, dual‐luciferase report; PLA, proximity ligation assay.

Article Snippet: The in situ PLA was performed on fixed GES‐1 cells following the manufacturer's protocol (DUO82049, DUO92008; Merck).

Techniques: Inhibition, In Vitro, Mutagenesis, Binding Assay, Electrophoretic Mobility Shift Assay, Autoradiography, Labeling, Sequencing, Luciferase, Activity Assay, Quantitative RT-PCR, Expressing, Negative Control, Staining, Western Blot, Targeted Gene Expression, Plasmid Preparation, Standard Deviation, Proximity Ligation Assay

Journal: BMC Cancer

Article Title: β-ionone synergizes with 5-Fluorouracil to inhibit gastric cancer progression through PAX6-mediated cell cycle arrest

doi: 10.1186/s12885-026-15540-2

Figure Lengend Snippet: BI and 5-FU exerted independent antiproliferative effects on gastric cancer cells. A MTT assay was used to detect the effects of BI and 5-FU at different concentrations and time on the viability of MKN45 cells, AGS cells, and NCI-N87 cells ( n = 5); ( B-C ) MTT assay was used to detect the effects of BI, 5-FU or their combination at different concentrations for 72 h on the viability of MKN45 cells and AGS cells and the synergistic effect of combination treatment was estimated by calculation of CI index using Compusyn software; ( D ) MTT assay was used to detect the effects of BI (144 µmol/L) and 5-FU (0.22 µmol/L) for 72 h on the viability of GES-1 cells and HS5 cells ( n = 5). Data are presented as mean ± S.D. of triplicates. Statistical analyses were performed using one-way ANOVA. Compared with the control group, * P < 0.05, ** P < 0.01. ns, not significant

Article Snippet: The gastric adenocarcinoma cell lines - MKN45 cells, AGS cells, NCI-N87 cells and human normal gastric gland GES-1 cells were purchased from Wuhan Procell Life Technology Co.,Ltd. (Wuhan, China).

Techniques: MTT Assay, Software, Control

Journal: Journal of Inflammation Research

Article Title: Wei-Mi-Shu, a Stomach-Harmonizing Herbal Prescription, Alleviates Chronic Gastritis with Liver‑Stomach Disharmony by Modulating IL-6/STAT3 Signaling

doi: 10.2147/JIR.S556234

Figure Lengend Snippet: WMSP promoted the viability of H. pylori -infected GES-1 cells and inhibited cellular inflammation through IL-6/STAT3 signaling pathway. ( A ) The morphology of GES-1 cells was observed by inverted microscope. ( B ) The viability of GES-1 cells infected with H. pylori was assessed using the Cell Counting Kit-8 (CCK-8) assay. ( C ) The concentrations of IL-6, CRP, TNF-α, and IL-1β in cell culture supernatants were measured by ELISA. ( D ) RT-qPCR were used to detect the mRNA of JAK1, JAK2, and STAT3 in GES-1 cells. ( E ) The expression levels of p-JAK1, JAK1, p-JAK2, JAK2, p-STAT3, and STAT3 in each treatment group were analyzed via Western blot. ( F ) The statistical results of relative protein expression levels. ( G ) The gene expression level of gp130 in cells was investigated using RT-qPCR. ( H ) The protein expression level of gp130 in cells was investigated using and Western blot. ( I ) The statistical results of the relative expression levels of gp130. ** P < 0.01 vs Control. ## P < 0.01 vs Model. & P < 0.05, && P < 0.01 vs WMSP.

Article Snippet: Total protein was extracted from 0.2 g of gastric tissue and GES-1 cells using RIPA lysate (Servicebio, Wuhan, China, #G2002).

Techniques: Infection, Inverted Microscopy, Cell Counting, CCK-8 Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Expressing, Western Blot, Gene Expression, Control