Establishment and characteristic analysis of a novel patient derived cell line of intrahepatic cholangiocarcinoma.

BACKGROUND: Intrahepatic cholangiocarcinoma (ICC) is a primary malignant tumor of the liver, second only to hepatocellular carcinoma. The scarcity of appropriate cell lines has impeded ICC-related research. This study aims to establish a novel patient-derived ICC cell line and systematically evaluate its biological characteristics, thereby providing a valuable cellular tool for ICC research. METHODS: A cell line named LPHC-6 of ICC was established using a patient-derived xenograft (PDX) tumor model. We comprehensively characterized this novel cell line through a series of techniques, including cell morphology observation, short tandem repeat (STR) analysis, karyotype analysis, cell proliferation, cell migration assays, western blot analysis, immunofluorescence staining, flow cytometry, gene mutation detection, spheroid formation assays, drug sensitivity testing, and xenograft tumor formation in nude mice. RESULTS: LPHC-6 cells exhibited typical characteristics of malignant epithelial cells, demonstrating robust proliferation and migration capabilities. These cells possessed a structurally complex triploid karyotype, which was confirmed to be free from cross-contamination with other human cell lines. Tumor biomarker analysis revealed positive expression of AFP and negative expression of GPC3. Additionally, LPHC-6 cells showed high expression levels of stem cell markers CD133 and CD44. The biliary epithelial cell marker CK19 was positive, whereas the hepatocyte marker Hep Par1 was negative. Genomic analysis identified an exon mutation in TP53, copy number loss in PTEN and RAD51, and copy number gain in NTRK1, PDCD1LG2, and VEGFA. Investigations confirmed that LPHC-6 cells exhibited excellent sphere formation ability and tumorigenic potential. Furthermore, LPHC-6 cells demonstrated sensitivity to Sorafenib, Lenvatinib, 5-fluorouracil, Oxaliplatin and Atezolizumab in both 2D and 3D culture system. CONCLUSION: A novel intrahepatic cholangiocarcinoma cell line LPHC-6 has been successfully established, which provides a powerful cell tool for the research related to intrahepatic cholangiocarcinoma.