Establishment and characterization of a highly metastatic hepatocellular carcinoma cell line

The prevalence of alcohol-related hepatocellular carcinoma (HCC) has been increasing during the last decade. Cancer research requires cell lines suitable for both <i>in vitro</i> and <i>in vivo</i> assays. However, there is a lack of cell lines with a high <i>in vivo</i> metastatic capacity for this HCC subtype. Herein, a new HCC cell line was established, named HCC-ZJ, using cells from a patient diagnosed with alcohol-related HCC. The karyotype of HCC-ZJ was 46, XY, del (p11.2). Whole-exome sequencing identified several genetic variations in HCC-Z that occur frequently in alcohol-associated HCC, such as mutations in <i>TERT</i>, <i>CTNNB1</i>, <i>ARID1A</i>, <i>CDKN2A</i>, <i>SMARCA2</i>, and <i>HGF</i>. Cell counting kit-8 assays, colony formation assays, and Transwell assays were performed to evaluate the proliferation, migration, and sensitivity to sorafenib and lenvatinib of HCC-Z <i>in vitro</i>. HCC-ZJ showed a robust proliferation rate, a weak foci-forming ability, a strong migration capacity, and a moderate invasion tendency <i>in vitro</i>. Finally, the tumorigenicity and metastatic capacity of HCC-Z were evaluated using a subcutaneous xenograft model, an orthotopic xenograft model, and a tail-veil injection model. HCCZJ exhibited strong tumorigenicity in the subcutaneous xenograft and orthotopic tumor models. Moreover, HCC-ZJ spontaneously formed pulmonary metastases in the orthotopic tumor model. In summary, a new HCC cell line derived from a patient with alcohol-related HCC was established, which showed a high metastatic capacity and could be applied for <i>in vitro</i> and <i>in vivo</i> experiments during pre-clinical research. <b>Highlights</b> • An alcohol-related HCC cell line, HCC-ZJ, was established • HCC-ZJ was applicable for <i>in vitro</i> functional experiment and gene editing • HCC-ZJ was applicable for <i>in vivo</i> tumor growth and spontaneous metastasis models