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. 2024 Sep 2;44(11):1231–1260. doi: 10.1002/cac2.12607

FIGURE 4.

FIGURE 4

SRSF10 positively regulated glycolysis in HCC cells. (A) GSEA analysis of differentially expressed gene from bulk RNA‐seq conducted on control and SRSF10‐knockdown Huh7 cells. (B) Fold change of glycolysis‐related genes in patients with high and low expression of SRSF10 in the TCGA‐LIHC dataset. (C) KEGG analysis of differentially expressed metabolite from untargeted energy metabolism sequencing conducted on control and shSrsf10 Hepa1‐6 cells. (D) Western blotting was performed in Huh7, PLC/PRF/5, and Hepa1‐6 cells. (E) Detection of lactate content in Huh7, PLC/PRF/5, and Hepa1‐6 cells (n = 3). (F‐G) ECAR of Huh7 cells and Hepa1‐6 cells (n = 5). (H) The expression SRSF10 mRNA in Huh7 and PLC/PRF/5 cells treated with LA, detected by qRT‐PCR analyses (n = 3). (I) SRSF10 mRNA levels in Huh7 and PLC/PRF/5 cells treated with 2‐DG, detected by qRT‐PCR analyses (n = 3). (J) Western blotting was performed in Huh7 and PLC/PRF/5 cells treated with LA. (K) Western blotting was performed in Huh7 and PLC/PRF/5 cells treated with 2‐DG. (L) H3K18la binding to the SRSF10 promoter was determined by chromatin immunoprecipitation‐RT‐PCR in Huh7 and PLC/PRF/5 cells (n = 3). (M) Schematic proposing a positive feedback loop between SRSF10, glycolysis and histone lactylation in HCC cells. Two‐tailed unpaired Student's t test (E‐I, L); ns, not significant; * P < 0.05, ** P < 0.01, *** P < 0.001. 2‐DG, 2‐Deoxy‐D‐glucose; DMSO, Dimethyl Sulfoxide; ECAR, Extra Cellular Acidification Rate; ENO1, enolase 1; ENO2, enolase 2; FDR, False Discovery Rate; GLUT1, Glucose transporter 1; GSEA, Gene Set Enrichment Analysis; HK1, hexokinase 1; HK2, hexokinase 2; HK3, hexokinase 3; IL2, interleukin 2; IL6, interleukin 6; LA, lactic acid; LDHA, lactate dehydrogenase A; LDHB, lactate dehydrogenase B; LIHC, Liver hepatocellular carcinoma; NES, Normalized enrichment score; PBS, Phosphate buffered saline; PGK1, phosphoglycerate kinase 1; PKM, pyruvate kinase M1/2; qRT‐PCR, Quantitative Real‐Time Reverse Transcription Polymerase Chain Reaction; RNA‐seq, RNA Sequencing; SRSF10, serine and arginine rich splicing factor 10; STAT3, signal transducer and activator of transcription 3; STAT5, signal transducer and activator of transcription 5; TCGA, The Cancer Genome Atlas.