在过去的十年中,酒精相关的肝细胞癌(HCC)的患病率一直在增加。癌症研究需要适用于体外和体内测定的细胞系。然而,这种HCC亚型缺乏具有高体内转移能力的细胞系。在这里,建立了一个新的肝癌细胞系,命名为HCC-ZJ,使用来自诊断为酒精相关HCC的患者的细胞。HCC-ZJ的核型为46,XY,del(p11.2).全外显子组测序确定了HCC-Z中的几种遗传变异,这些变异在酒精相关HCC中频繁发生,例如TERT中的突变,CTNNB1,ARID1A,CDKN2A,SMARCA2和HGF。细胞计数试剂盒-8测定,集落形成试验,进行Transwell分析以评估增殖,迁移,肝癌-Z的体外对索拉非尼和乐伐替尼的敏感性。HCC-ZJ显示出强劲的增殖率,聚焦形成能力弱,强大的迁移能力,在体外有中等的侵袭倾向。最后,使用皮下异种移植模型评估HCC-Z的致瘤性和转移能力,原位异种移植模型,和尾部面纱注射模型。HCCZJ在皮下异种移植和原位肿瘤模型中表现出强烈的致瘤性。此外,HCC-ZJ在原位肿瘤模型中自发形成肺转移。总之,建立了一个来自酒精相关HCC患者的新HCC细胞系,显示出高转移能力,可用于临床前研究期间的体外和体内实验。亮点•酒精相关的肝癌细胞系,HCC-ZJ,•HCC-ZJ适用于体外功能实验和基因编辑•HCC-ZJ适用于体内肿瘤生长和自发转移模型。
The prevalence of alcohol-related hepatocellular carcinoma (HCC) has been increasing during the last decade. Cancer research requires cell lines suitable for both in vitro and in vivo assays. However, there is a lack of cell lines with a high in vivo 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 TERT, CTNNB1, ARID1A, CDKN2A, SMARCA2, and HGF. 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 in vitro. HCC-ZJ showed a robust proliferation rate, a weak foci-forming ability, a strong migration capacity, and a moderate invasion tendency in vitro. 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 in vitro and in vivo experiments during pre-clinical research.Highlights• An alcohol-related HCC cell line, HCC-ZJ, was established• HCC-ZJ was applicable for in vitro functional experiment and gene editing• HCC-ZJ was applicable for in vivo tumor growth and spontaneous metastasis models.