Establishment and characterization of a new human gallbladder cancer cell line, OCUG-2

Figure 1 Computed tomography scan and histopathological findings of a patient with gall bladder cancer. A: Computed tomography (CT) findings of the primary tumor. CT scan image of the transverse and coronal planes display gallbladder carcinoma (arrows); B: Hematoxylin and eosin staining of the metastatic peritoneal tumor of patients with gallbladder cancer. The tumor showed poorly differentiated growth with moderate stromal cells.

Figure 2 Morphologic findings of OCUG-2 cells. A: Photomicrography of living OCUG-2 cells; B: Growth curve of OCUG-2 cells; C: Hematoxylin and eosin (H&E)-stained section of subcutaneous tumors induced by OCUG-2 cell injection in nude mice. Tumor histology was consistent with that of the original primary tumor, as shown by H&E staining. Scale bar, 100 μm.

Figure 3 Expression of growth factor receptors and growth kinetics. A: mRNA expression of growth factor receptors EGFR, HER2, ERBB3, ERBB4, PDGFR-α, PDGFR-β, insulin-like growth factor 1 receptor, FMS-like tyrosine kinase, FGFR1, FGFR2, FGFR3, FGFR4, VEGFR1, VEGFR2, VEGFR3, c-Kit, and c-Met; B: Growth curves of OCUG-2 cells treated with panitumumab, cetuximab, bevacizumab, ramucirumab, CP673451, picropodophyllin, SU11274, infigratinib, FGF401, afatinib, sapitinib, and trastuzumab. Growth of OCUG-2 cells was significantly decreased by picropodophyllotoxin.

Figure 4 Invasion ability of OCUG-2 cells and insulin-like growth factor 1 receptor staining of gall bladder tumors. A: Effects of various targeted drugs on the invasion ability of OCUG-2 cells. Invasion ratio of OCUG-2 cells post-treatment compared to controls. Photograph (200 ×) of OCUG-2 invasion cells treated with control or SU11274; B: Immunohistochemical staining of insulin-like growth factor 1 receptor (IGF1R). IGF1R expression was found on cancer cell membrane. Scale bar, 100 μm. Kaplan-Meier survival analysis comparing overall survival and disease-free survival in IGF1R-positive and IGF1R-negative gallbladder cancer patients. ^aP < 0.05, ^bP < 0.01, and ^cP < 0.001.

Figure 5 RNA-sequencing characterization of OCUG-2 cells. A: Circular graph depicting single nucleotide polymorphisms and indel variations in the OCUG-2 genome; B: Mutation spectrum of single nucleotide variants in the OCUG-2 genome; C: Gene Ontology analysis and the significantly enriched Kyoto Encyclopedia of Genes and Genomes pathways of the top 20 mutation genes selected based on q-values. GO: Gene Ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes.