Fig 2: S100A10 promotes aerobic glycolysis in GC. (A–H) The influence of S100A10 overexpression or knockdown on glucose consumption (A,B), lactate production (C,D), relative LDH activity (E,F), and ATP production (G,H) in GC cells. (I–L) The influence of S100A10 knockdown on the extracellular acidification ratio (ECAR) and oxygen consumption ratio (OCR) in GC cells. *P < 0.05, **P < 0.01, ***P < 0.001.

Fig 3: S100A10 is closely correlated with glycolysis. (A) S100A10 showed the highest expression in the metabolic subtype of GC. (B–D) RNA-seq data of stomach adenocarcinoma samples were downloaded from the TCGA database. These samples were defined as S100A10-high and S100A10-low groups based on S100A10 expression. Gene set enrichment analysis (GSEA) was then performed and revealed that S100A10 was closely related to insulin signaling pathway, oxidative phosphorylation and mTOR signaling pathway. NES, normalized enrichment score; NOM, nominal; FDR, false discovery rate. (E–K) Correlation analysis between S100A10 and glycolytic transporters or enzymes in GC, including SLC2A1 (GLUT1), SLC2A4 (GLUT4), HK2, IDH1, LDHA, PFKFB3, and PKM. (L–O) The mRNA expression levels of glycolytic transporters or enzymes were assessed by RT-qPCR after S100A10 overexpression or knockdown. *P < 0.05, **P < 0.01, ***P < 0.001.

Fig 4: S100A10 is succinylated at lysine 47 in GC. A, Immunoprecipitation analysis of S100A10 succinylation in GC tissues. B, Identification of succinylated S100A10 peptides by mass spectrometry of GC tissues. C, Protein amino acid sequence alignment of S100A10 surrounding K47 from various organisms. K47 is shown in red. The peptide sequence used for preparing the antibody is indicated. D, K47 is the primary succinylation site of S100A10. The indicated plasmids were transfected into MGC-803 cells and proteins were immunoprecipitated, followed by western blotting for succinylation analysis. E, Western blot analysis of S100A10 K47succ expression in various cell lines. F, Western blot analysis of the relative levels of S100A10 K47succ expression in seven pairs of GC and adjacent non-cancerous tissues. G, Immunohistochemistry of S100A10 K47succ in adjacent normal tissues, GC tissues, normal lymph nodes and metastatic lymph nodes of GC patients. Results are derived from ten tissue slides. Data are presented as mean ± SEM; *P < 0.05

Fig 5: Evidence that S100A10 functions as an oncogene in GC. (A) The mRNA expression of S100A10 in different human cancers and corresponding normal tissues from the Oncomine database. The number in the colored cell is the number of datasets meeting the thresholds. Cell color is determined by the gene rank. The color intensity (red or blue) is directly proportional to the significance level of upregulation or downregulation, respectively. (B,C) S100A10 expression in cancers. Expression level of S100A10 across 33 TCGA tumors compared to TCGA normal and GTEx data using the GEPIA (Gene Expression Profiling Interactive Analysis) webserver. An obvious upregulation of this gene was evident in 12 cancers. For each TCGA tumor (red), its matched normal and GTEx data (green) are given. The abbreviations denote T: tumor; N: normal; n: number. Y-axis: transcript per million [log2 (TPM + 1)]. X-axis: number of tumor and normal samples. ACC: adrenocortical carcinoma; BLCA: bladder urothelial carcinoma; BRCA: breast invasive carcinoma; CESC: cervical squamous cell carcinoma and endocervical adenocarcinoma; CHOL: cholangiocarcinoma; COAD: colon adenocarcinoma; DLBC: lymphoid neoplasm diffuse large B-cell lymphoma; ESCA: esophageal carcinoma; GBM: glioblastoma multiforme; HNSC: head and neck squamous cell carcinoma; KICH: kidney chromophobe; KIRC: kidney renal clear cell carcinoma; KIRP: kidney renal papillary cell carcinoma; LAML: acute myeloid leukemia; LGG: brain lower grade glioma; LIHC: liver hepatocellular carcinoma; LUAD: lung adenocarcinoma; LUSC: lung squamous cell carcinoma; MESO: mesothelioma; OV: ovarian serous cystadenocarcinoma; PAAD: pancreatic adenocarcinoma; PCPG: pheochromocytoma and paraganglioma; PRAD: prostate adenocarcinoma; READ: rectum adenocarcinoma; SARC: sarcoma; SKCM: skin cutaneous melanoma; STAD: stomach adenocarcinoma; TGCT: testicular germ cell tumors; THCA: thyroid carcinoma; THYM: thymoma; UCEC: uterine corpus endometrial carcinoma; UCS: uterine carcinosarcoma; and UVM: uveal melanoma. (D) The expression level of S100A10 in GC and normal tissues using data from the TCGA database. (E) The expression of S100A10 in GC based on individual stages using the UALCAN database. (F) The expression of S100A10 in different GC subtypes compared to normal individuals derived from the Oncomine database. (G) S100A10 expression levels were associated with poor survival, as analyzed by the online program Kaplan-Meier Plotter. *P < 0.05, **P < 0.01, ***P < 0.001.