Establishment and characterization of a novel highly aggressive hilar cholangiocarcinoma cell line, CBC3T-1

Abstract Background Cholangiocarcinoma (CCA) is a group of malignant heterogeneous cancers arising from the biliary tree characterized by insidious onset, poor prognosis and high recurrence rate. Immortalized cancer cell lines are the best and easiest models for in vitro and in vivo cancer research. Methods CBC3T-1 was originated from a patient with hilar CCA from China and identified by STR and chromosome karyotype analysis. The malignant phenotype was characterized by wound healing assay, transwell migration, Matrigel invasion assay, clone formation assay and in vivo xenograft mouse models. Screening of sensitive clinical anticancer drugs by CCK-8 assay. The cell line was characterized at the genetic level using transcriptome sequencing and whole exome sequencing. Results CBC3T-1 was cultured for over 60 passages. Thorough analysis showed that CBC3T-1 cells share characteristics similar to original tumor cells from patients with cholangiocarcinoma and display a stable phenotype, including features of epithelial origin, stem cell-like properties as well as a high invasive and migratory capability and tumorigenicity in mice. Furthermore, this cell line showed the best sensitivity to paclitaxel, followed by gemcitabine. Sequencing results show that cancer-associated pathways and somatic mutations play a dominant role in the development of CCA. Conclusions We established and characterized a novel highly aggressive hilar CCA cell line, CBC3T-1, which provides a new model for studying the progression of hilar CCA as well as developing new therapeutic approaches, enriching the heterogeneous in vitro models of CCA.