Abstract 1662: Orthotopic model of hepatocellular carcinoma (HCC) using Huh-7 and HepG2: Application of micro-CT in detection of cancer progression
Authors: Zdrojewska Justyna , Suominen Mari , Katoozi Shirin , Fagerlund Katja , Rissanen Jukka , Malerbakken-Bjerke Roger , Karlsson Jenny 
Published: June 2022

Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer and was listed as the third cause of death by cancer in 2020 (WHO). Hepatocarcinogenesis is a multistep process and to date it is still poorly understood. HCC occurs almost entirely in patients with underlying chronic liver disease and cirrhosis. Pre-clinical animal models play an important role in unveiling cancer biology which can facilitate development and testing of new antineoplastic drugs. The aim of this study was to establish an orthotopic model of HCC utilizing two cell lines Huh-7 and HepG2, which could be effectively used to study efficacy of new potential treatments. In a follow up study, the correlation of tumor marker alpha-fetoprotein (AFP) and tumor growth was investigated. AFP was measured using Quantikine® ELISA kit. For the intrahepatic (ih) inoculation cancer cells were injected into the left lobe of the liver. For the intrasplenic (is) inoculations of HCC cells, the spleen was exposed, and cells were injected into spleen parenchyma along the long axis of the spleen. Afterwards spleen was removed to avoid intrasplenic tumor growth. The animals were weighed two times a week throughout the study and appearance of any clinical signs was closely monitored. The animals were terminated 70 days after cancer cell implantation. The mice did not present with any acute symptoms before the end stage disease. However, prior sacrifice some swelling and darkening of the abdominal area especially in the ih-model was observed. To visualize liver ExiTron Nano 6000 nanoparticle contrast agent was used. The contrast agent was administered intravenously once during the study, prior the first imaging. Disease progression was monitored using Bruker SkyScan 1276 High-Resolution Micro-CT Scanner. The in vivo measurements were performed using one field of view (FOV) and an image pixel size of 18 μm. Each in vivo scanning was completed within 12 minutes and the estimated radiation dose was kept under 500 mGy at each imaging session. The first tumours were detected by µCT between 2-3 weeks depending more on the cancer cell implantation route than the cell line used. The take rate in the liver was 85% in the ih-model, and less than 50% in the is-model. The intrahepatic inoculation led to more uniform and measurable tumor nodules observed in majority of operated animals, whereas the intrasplenic tumors were dispersed throughout with multiple nodules. In addition, a tight correlation between tumor volume and amount of alpha-fetoprotein detected from serum samples was found. In conclusion, an orthotopic model using intrahepatic approach of cancer cells implantation was established. Application of µCT enabled following tumor growth and recognition of cancer spread in the abdomen. Altogether this provides a promising tool contributing to the development of novel therapies targeting HCC in vivo.

Citation Format: Justyna Zdrojewska, Mari Suominen, Shirin Katoozi, Katja Fagerlund, Jukka Rissanen, Roger Malerbakken-Bjerke, Jenny Karlsson. Orthotopic model of hepatocellular carcinoma (HCC) using Huh-7 and HepG2: Application of micro-CT in detection of cancer progression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1662.