Journal: Translational Cancer Research

Article Title: SLC25A3 promotes hepatocellular carcinoma progression by inducing mitochondrial dysfunction and chromatin remodeling

doi: 10.21037/tcr-2025-1946

Figure Lengend Snippet: SLC25A3 in highly expressed in HCC and correlates with poor prognosis. (A) Expression of mitochondrial-related genes in HCC. (B) Differential expression of SLC25A3 in HCC tissues and normal liver tissues at the mRNA level. (C) Differential expression of SLC25A3 in HCC tissues and normal liver tissues at different pathological stages. (D) DNA methylation status of SLC25A3 in HCC tissues and normal liver tissues. (E) DNA methylation status of SLC25A3 in HCC tissues and normal liver tissues at different pathological stages. (F) Kaplan-Meier survival curves for high/low expression groups of SLC25A3 in HCC patients based on TCGA data. ***, P<0.001. CI, confidence interval; HCC, hepatocellular carcinoma; HR, hazard ratio; TCGA, The Cancer Genome Atlas; TPM, transcripts per million.

Article Snippet: Briefly, the tissue sections were incubated overnight at 4 °C with anti-SLC25A3 antibody (1:100, #10420-1-AP, Proteintech, Chicago, USA) on the first day.

Techniques: Expressing, Quantitative Proteomics, DNA Methylation Assay

Journal: Translational Cancer Research

Article Title: SLC25A3 promotes hepatocellular carcinoma progression by inducing mitochondrial dysfunction and chromatin remodeling

doi: 10.21037/tcr-2025-1946

Figure Lengend Snippet: Investigation of SLC25A3 expression in HCC based on clinical samples and cell lines. (A) Western blot detection of SLC25A3 expression at the protein level in HCC and adjacent liver tissues (n=20) (T: tumor; P: peri-tumor). (B) Immunohistochemical analysis of SLC25A3 expression and localization in HCC and adjacent liver tissues (scale bar =100 µm). (C) Quantitative analysis of IHC results (n=50). (D) Western blot detection of SLC25A3 expression at the protein level in HCC cell lines (Huh7, HCC-LM3, MHCC-97H, PLC/PRF/5, HepG2) and normal liver cell line (L02). ***, P<0.001. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HCC, hepatocellular carcinoma; IHC, immunohistochemistry.

Article Snippet: Briefly, the tissue sections were incubated overnight at 4 °C with anti-SLC25A3 antibody (1:100, #10420-1-AP, Proteintech, Chicago, USA) on the first day.

Techniques: Expressing, Western Blot, Immunohistochemical staining, Immunohistochemistry

Journal: Translational Cancer Research

Article Title: SLC25A3 promotes hepatocellular carcinoma progression by inducing mitochondrial dysfunction and chromatin remodeling

doi: 10.21037/tcr-2025-1946

Figure Lengend Snippet: SLC25A3 knockout impaired hepatocarcinogenesis by inducing morphological changes in mitochondria. (A) Genotyping of SLC25A3-KO and WT mice using DNA extracted from tail tissue (lanes 4/5 are WT mice, others are SLC25A3-KO mice). (B) The efficiency of SLC25A3 knockout in mice was validated by western blot. (C) Macroscopic photography of DEN-induced HCC mice models. (D) Quantitative analysis of liver-to-body weight ratio, tumor number, maximum tumor diameter, and blood biochemical markers (ALT, AST, TBIL, DBIL) in both groups. (E) Scanning electron microscopy of liver ultrastructure in both groups of mice (m, mitochondria; scale bar =1 µm). *, P<0.05; **, P<0.01; ns, not significant. ALT, alanine aminotransferase; AST, aspartate aminotransferase; DBIL, direct bilirubin; DEN, diethylnitrosamine; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; KO, knockout; TBIL, total bilirubin; WT, wild-type.

Article Snippet: Briefly, the tissue sections were incubated overnight at 4 °C with anti-SLC25A3 antibody (1:100, #10420-1-AP, Proteintech, Chicago, USA) on the first day.

Techniques: Knock-Out, Western Blot, Electron Microscopy

Journal: Translational Cancer Research

Article Title: SLC25A3 promotes hepatocellular carcinoma progression by inducing mitochondrial dysfunction and chromatin remodeling

doi: 10.21037/tcr-2025-1946

Figure Lengend Snippet: SLC25A3 promoting hepatocarcinogenesis by inducing mitochondrial dysfunction. (A) Efficiency of plasmid overexpression and knockdown in HCC cell lines, assessed by western blot. (B) Cell viability assessed by CCK-8 assay. (C) HCC-LM3 cells stained with crystal violet. (D) Huh7 cells stained with crystal violet. (E,F) Mitochondrial oxidative phosphorylation levels measured by ATP content and basal OCR. (G) Mitochondrial membrane potential, assessed by JC-1 staining, represented as the relative aggregate-to-monomer (red/green) fluorescence intensity ratio (100×). **, P<0.01; ***, P<0.001; ns, not significant. ATP, adenosine triphosphate; CCK-8, Cell Counting Kit8; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; HCC, hepatocellular carcinoma; KD, knockdown; Mock, mock control; NC, negative control; OCR, oxygen consumption rate; OE, overexpression.

Article Snippet: Briefly, the tissue sections were incubated overnight at 4 °C with anti-SLC25A3 antibody (1:100, #10420-1-AP, Proteintech, Chicago, USA) on the first day.

Techniques: Plasmid Preparation, Over Expression, Knockdown, Western Blot, CCK-8 Assay, Staining, Phospho-proteomics, Membrane, Fluorescence, Cell Counting, Control, Negative Control

Journal: Translational Cancer Research

Article Title: SLC25A3 promotes hepatocellular carcinoma progression by inducing mitochondrial dysfunction and chromatin remodeling

doi: 10.21037/tcr-2025-1946

Figure Lengend Snippet: Multi-omics sequencing analysis to investigate the impact of SLC25A3 on epigenetic modifications and gene expression. (A) GSEA of RNA-seq results from SLC25A3-KO and WT mouse livers. (B) Differential analysis of mRNA-seq results, with a volcano plot showing DEGs. (C) KEGG functional analysis of DEGs from RNA-seq. (D) ATAC-seq analysis of chromatin conformation changes (SLC25A3-KO, KO group; WT, NC group). (E) KEGG pathway analysis of gene regions exhibiting chromatin conformational changes identified in ATAC-seq. (F) Joint analysis of ATAC-seq and RNA-seq results to identify genes with tight chromatin conformation and decreased expression in the SLC25A3-KO group. (G) KEGG functional analysis of genes identified in the joint analysis. ATAC-seq, Assay for Transposase-Accessible Chromatin using sequencing; DEGs, differentially expressed genes; FC, fold change; GSEA, gene set enrichment analysis; KEGG, Kyoto Encyclopedia of Genes and Genomes; KO, knockout; NC, negative control; RNA-seq, RNA sequencing; WT, wild-type.

Article Snippet: Briefly, the tissue sections were incubated overnight at 4 °C with anti-SLC25A3 antibody (1:100, #10420-1-AP, Proteintech, Chicago, USA) on the first day.

Techniques: Biomarker Discovery, Sequencing, Gene Expression, RNA Sequencing, Functional Assay, Expressing, Knock-Out, Negative Control