Targeted gene sequencing and bioinformatics analysis of patients with gallbladder neuroendocrine carcinoma: A case report

Figure 1 Abdominal contrast-enhanced computed tomography scan reveals a 7 cm mass in liver segments IVb and V, with localized thickening of the gallbladder fundus wall and early-stage enhancement followed by a prolonged contrast effect. A: Arterial phase showed inhomogeneous lesion enhancement; B: The portal venous phase showed a prolonged contrast effect in the mass; C: The delayed phase showed progressive enhancement of the mass; D: The mass invades liver segments IVb and V.

Figure 2 Preoperative magnetic resonance imaging examination of the reported case. A and B: The gallbladder lesion appears slightly hyperintense on T2-weighted imaging (A) and hyperintense on diffusion-weighted imaging (B).

Figure 3 Illustration of the surgical procedure. A: A radical cholecystectomy was performed; B: The gross tumor specimen was displayed; C: Macroscopic analysis revealed a greyish-yellow globular lesion; D: Multiple yellowish stones were identified.

Figure 4 Pathological analysis results. A: Hematoxylin and eosin staining revealed a nest-like arrangement of tumor cells exhibiting marked cellular heterogeneity and visible necrosis; B–D: Immunohistochemical analyses showed positivity for chromogranin A, Ki-67, and synaptophysin, respectively.

Figure 5 PD-L1 and tumor mutation burden. A and B: Targeted gene sequencing revealed no expression of PD-L1 in gallbladder neuroendocrine carcinoma cells; C and D: Buffer solutions and isotype-matched monoclonal antibodies were used as controls to confirm the specificity of primary antibody binding; E: Targeted gene sequencing also helped quantify the tumor mutation burden. TMB: Tumor mutation burden.

Figure 6 Interaction network analyses. A and B: The protein-protein interaction network was constructed using 12 mutated genes; C and D: Protein-protein interaction enrichment analysis was performed to explore functional relationships between these 12 mutated genes.

Figure 7 Significantly enriched Gene Ontology terms and Kyoto Encyclopedia of Genes and Genomes pathways for mutated genes. A-D: Gene Ontology analysis of biological processes (A), cellular components (B), molecular function (C), and Kyoto Encyclopedia of Genes and Genomes enrichment analysis (D); E and F: The Metascape database yielded similar results for the mutated genes. GO: Gene Ontology; BP: Biological processes; CC: Cellular components; MF: Molecular function; KEGG: Kyoto Encyclopedia of Genes and Genomes.

Figure 8 Gene mutations in gallbladder carcinomas. A: A summary of alterations in the 12 queried gene mutations from the cBioPortal database; B: Detailed summary of alterations in the 12 mutated genes; C: The correlation between KEAP1 expression and the sensitivity to GDSC drugs (top 30) in pan-cancer analysis.