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

Figure 7 Cdkn1c knockdown attenuates pristimerin’s regulation of glycolytic reprogramming and amelioration of spasmolytic polypeptide-expressing metaplasia. A: Representative images of normal gastric organoids, N-methyl-N’-nitro-N-nitrosoguanidine/Helicobacter pylori-induced spasmolytic polypeptide-expressing metaplasia (SPEM) gastric organoids, and those following Cdkn1c knockdown or pristimerin intervention, along with quantitative analyses of organoid number and diameter. Scale bar, 100 μm; B: Effects of Cdkn1c knockdown or pristimerin treatment on p57 expression levels in gastric organoids. Scale bar, 10 μm; C-E: Cdkn1c knockdown promotes cell proliferation [reverse transcription quantitative polymerase chain reaction (RT-qPCR); Western blot, β-actin-normalized relative expression vs control] and enhances the expression of key glycolytic regulators (RT-qPCR; Western blot), stem cell markers (immunofluorescence: Scale bar, 10 μm), and SPEM-specific markers (RT-qPCR; Western blot) in SPEM gastric organoids, while suppressing gastric intrinsic factor (RT-qPCR; Western blot) expression. It also attenuates the regulatory effects of pristimerin on these phenotypes. ^aP < 0.05. ^bP < 0.01. ^cP < 0.001. ^dP < 0.0001. Values represent the mean ± SD (n = 5). Cdkn1c-KD: Cdkn1c-knockdown spasmolytic polypeptide-expressing metaplasia organoids; DAPI: 4’,6-diamidino-2-phenylindole; GIF: Gastric intrinsic factor; Pri: Pristimerin; IOD: Integrated optical density; HIF-1α: Hypoxia-inducible factor-1α; LDHA: Lactate dehydrogenase A; PKM2: Pyruvate kinase M2; 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.