Establishment and characterization of a highly metastatic hepatocellular carcinoma cell line

ABSTRACT

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

KEYWORDS:

• Hepatocellular carcinoma
• cell line
• xenograft
• primary culture
• metastasis

Disclosure statement

No potential conflict of interest was reported by the author(s).

Author contribution

Conceptualization, Zhenhua Hu and Yiting Qiao; methodology, Zhenhua Hu and Yiting Qiao; software, Jiacheng Huang and Mengqing Sun; validation, Jiacheng Huang, Mengqing Sun, Menglan Wang, Anning Yu and Chiwen Bu; formal analysis, Jiacheng Huang, Mengqing Sun, Menglan Wang, Anning Yu and Chiwen Bu; investigation, Huilin Zheng; resources, Jie Zhou and Yu Zhang; data curation, Jiacheng Huang and Huilin Zheng; writing – original draft preparation, Jiacheng Huang and Mengqing Sun; writing – review and editing, Jiacheng Huang, Mengqing Sun, Menglan Wang, Anning Yu and Chiwen Bu; visualization, Jiacheng Huang, Mengqing Sun; supervision, Zhenhua Hu and Yiting Qiao; project administration, Zhenhua Hu and Yiting Qiao; funding acquisition, Zhenhua Hu, Yiting Qiao and Chiwen Bu. All authors have read and agreed to the published version of the manuscript.

Data availability statement

The data presented in this study are available on request from the corresponding author. The data are not publicly available due to the patient’s privacy.

Informed consent statement

Informed consent was obtained from all subjects involved in the study. Written informed consent has been obtained from the patient(s) to publish this paper

Institutional review board statement

The study was conducted in accordance with the Declaration of Helsinki, and approved by the Clinical Research Ethics Committee of the First Affiliated Hospital, Zhejiang University School of Medicine (Reference number: IIT20200623A). The animal study protocol was approved by the Animal Experiment Ethical Inspection of the First Affiliated Hospital, Zhejiang University School of Medicine (Reference number: 2021907). Our animal study adhered to the Animal Research: Reporting of In Vivo Experiments (ARRIVE) guideline.

Supplementary material

Supplemental data for this article can be accessed online at https://doi.org/10.1080/21655979.2023.2296775

Additional information

Funding

This work was supported by the Fundamental Research Funds for the Central Universities (226-2023-00107); Research Project of Jinan Microecological Biomedicine Shandong Laboratory (JNL-2022029C); Natural Science Foundation of Zhejiang Province (LY21H160025); National Key Research and Development Program of China (2022YFA1104600); and Research Development Fund of Kangda College of Nanjing Medical University (KD2022KYCXTD013).