Exploring the potential function of high expression of ANAPC1 in regulating ubiquitination in hepatocellular carcinoma

Figure 1 The main analysis flow chart of this study. HCC: Hepatocellular carcinoma; mRNA: Messenger RNA; scRNA-seq: Single-cell RNA sequencing; SROC: Synthetic receiver operating characteristic; UMAP: Uniform manifold approximation and projection; NK: Natural killer; MIF: Migration inhibitory factor; MK: Midkine; VISFATIN: Visfatin; SPP1: Secreted phosphoprotein 1; PARs: Protease-activated receptors; ANGPTL: Angiopoietin-like protein; GALECTIN: Galactoside-binding lectin; CXCL: Chemokine (C-X-C motif) ligand; COMPLEMENT: Complement system proteins; PTN: Pleiotrophin; CCL: Chemokine (C-C motif) ligand; VEGF: Vascular endothelial growth factor; CALCR: Calcitonin receptor; ANNEXIN: Annexin; GDF: Growth differentiation factor; GAS: Growth arrest-specific 6; PROS: Protein S; ANGPT: Angiopoietin; PSAP: Prosaposin; BAFF: B cell activating factor; EGF: Epidermal growth factor; PDGF: Platelet-derived growth factor; IL16: Interleukin 16; FGF: Fibroblast growth factor; PERIOSTIN: Periostin.

Figure 2 Comprehensive analysis of ANAPC1 messenger RNA expression in hepatocellular carcinoma. A: Forest plot of integrated ANAPC1 messenger RNA (mRNA) expression from 38 platforms; B: Summary receiver operating characteristic curve; C: Begg’s test; D: Egger’s test. Calculated using the integration method, a higher area under the curve indicates a more pronounced difference in ANAPC1 mRNA expression between non-hepatocellular carcinoma and hepatocellular carcinoma tissues. CI: Confidence interval; MTAB: Matrix table; GPL: Gene platform; ICGC: International cancer genome consortium; TCGA: The cancer genome atlas; HCC: Hepatocellular carcinoma; SENS: Sensitivity; SPEC: Specificity; SROC: Synthetic receiver operating characteristic; AUC: Area under the curve.

Figure 3 Immunohistochemistry staining of non-hepatocellular carcinoma and hepatocellular carcinoma samples. A-E: Hepatocellular carcinoma samples; F-J: Non-hepatocellular carcinoma samples.

Figure 4 Comprehensive analysis of ANAPC1 protein expression. A: Forest plot of ANAPC1 protein comprehensive expression; B: Summary receiver operating characteristic curve; C: Begg’s test; D: Egger’s test. Calculated using the integration method, a higher area under the curve indicates a more distinct difference in ANAPC1 protein expression between non-hepatocellular carcinoma and hepatocellular carcinoma tissues. CI: Confidence interval; SENS: Sensitivity; SPEC: Specificity; SROC: Synthetic receiver operating characteristic; AUC: Area under the curve.

Figure 5 ANAPC1 expression profiling in single-cell RNA sequencing. A: Sample distribution plot from GSE14694; B: Cell type distribution plot; C: ANAPC1 expression levels across different cell types; D: Volcano plot. UMAP: Uniform manifold approximation and projection; HCC: Hepatocellular carcinoma; NK: Natural killer.

Figure 6 Preliminary exploration of the molecular function of ANAPC1. A: GeneMANIA analysis; B: Gene set enrichment analysis. NES: Normalized enrichment score; Pd-1: Programmed cell death protein 1; Pd-L1: Programmed death-ligand 1.

Figure 7 Enrichment analysis of highly expressed positive correlation genes related to ANAPC1. A: Gene ontology; B: Kyoto encyclopedia of genes and genomes; C: Reactome. rRNA: Ribosomal RNA; ncRNA: Non-coding RNA; ATP: Adenosine triphosphate; HIV: Human immunodeficiency virus; NEP/NS2: Nuclear export protein/non-structural protein 2.

Figure 8 Analysis of ANAPC1 metabolic levels in single-cell RNA sequencing. A: Expression levels of ANAPC1 in various clusters of malignant hepatocytes; B: Cluster distribution map of high and low ANAPC1 expression groups; C: Metabolic level analysis of high and low ANAPC1 expression groups. UMAP: Uniform manifold approximation and projection; TCA: Tricarboxylic acid cycle.

Figure 9 Regulation of high ANAPC1 expression in cell communication, particularly in the migration inhibitory factor pathway. A: Weights/strengths of cell-cell interactions; B: Signaling roles of different cell clusters in the Midkine signaling pathway; C: Heatmap of cell types involved in the migration inhibitory factor pathway; D: Distribution of signal transduction genes participating in the Midkine signaling pathway. NK: Natural killer; MIF: Migration inhibitory factor; CXCR: C-X-C chemokine receptor; CD: Cluster of differentiation.

Figure 10  Relationship between ANAPC1 expression levels and tyrosine kinase inhibitors drug effects. A: Sorafenib; B: Dasatinib; C: Ibrutinib; D: Lapatinib; E: Nilotinib; F: Afatinib. Ic50: Half maximal inhibitory concentration.