Pristimerin ameliorates spasmolytic polypeptide-expressing metaplasia by modulating Cdkn1c (p57)-mediated glycolytic reprogramming

Figure 5 Pristimerin ameliorates tamoxifen-induced spasmolytic polypeptide-expressing metaplasia phenotype in human gastric epithelial cells via suppression of glycolysis. A: Representative images of tamoxifen-induced human gastric epithelial (GES-1) cells following pristimerin or 2-deoxy-D-glucose (2-DG) intervention. Scale bar, 100 μm; B: Expression of key glycolytic regulators in GES-1 cells following pristimerin or 2-DG intervention; C: Quantitative analysis of pyruvic acid and lactate production in GES-1 cells; D: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay for GES-1 cell viability; E: 5-Ethynyl-2’-deoxyuridine assay for detecting proliferation activity in GES-1 cells. Scale bar, 20 μm; F: Inhibitory effects of pristimerin or 2-DG on spasmolytic polypeptide-expressing metaplasia-specific marker expression in GES-1 cells (Western blot; β-actin-normalized relative expression vs control); G: Expression of stem cell markers in GES-1 cells following pristimerin or 2-DG treatment. Scale bar, 20 μm. ^aP < 0.05. ^bP < 0.01. ^cP < 0.001. ^dP < 0.0001. Values represent the mean ± SD (n = 5). 2-DG: 2-deoxy-D-glucose; Pri: Pristimerin; HIF-1α: Hypoxia-inducible factor-1α; LDHA: Lactate dehydrogenase A; PKM2: Pyruvate kinase M2; TAM: Tamoxifen; DAPI: 4’,6-diamidino-2-phenylindole; EdU: 5-Ethynyl-2’-deoxyuridine; MUC6: Mucin 6; TFF2: Trefoil factor 2; WFDC2: Whey-acidic-protein four-disulfide core domain protein 2; Lgr5: Leucine-rich repeat-containing G-protein coupled receptor 5; Troy: Tumor necrosis factor receptor superfamily member 19; IOD: Integrated optical density.