In vivo analysis of FBXO45-mediated fibrosis and liver tumorigenesis in a chemically induced mouse model of hepatocellular carcinoma

• Published in: STAR protocols Vol. 4; no. 1; p. 102124
• Main Authors: Zhang, Jie, Lin, Xiao-Tong, Fang, Lei, Xie, Chuan-Ming
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 17.03.2023; Elsevier
• Subjects: Cancer; Health Sciences; Microscopy; Model Organisms; Molecular Biology; Protocol; Model Organisms; Microscopy; Molecular Biology; Cancer; Health Sciences
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2023.102124

FBXO45, an E3 ubiquitin ligase highly expressed in liver tumors, is positively correlated with poor survival of hepatocellular carcinogenesis (HCC) patients, but whether FBXO45 drives HCC tumorigenesis remains largely unclear. Here, we describe a protocol that shortens the observation period for HCC tumorigenesis to assess the effects of FBXO45 in a DEN/CCl4-induced HCC mouse model. We describe steps for chemical induction of HCC in FBXO45-overexpressing mice, followed by tissue collection and pathology assessment via quantitative real-time PCR, histology, and immunohistochemistry. For complete details on the use and execution of this protocol, please refer to Lin et al. (2021).1 [Display omitted] •Protocol to chemically induce HCC in FBXO45-overexpressing mice•Procedure for the combined treatment of mice with DEN/CCl4 to induce HCC•Assessment of cell morphology and gene and protein expression in liver tissues•Applicable to assess other genes of interest in chemically induced HCC mouse models Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. FBXO45, an E3 ubiquitin ligase highly expressed in liver tumors, is positively correlated with poor survival of hepatocellular carcinogenesis (HCC) patients, but whether FBXO45 drives HCC tumorigenesis remains largely unclear. Here, we describe a protocol that shortens the observation period for HCC tumorigenesis to assess the effects of FBXO45 in a DEN/CCl4-induced HCC mouse model. We describe steps for chemical induction of HCC in FBXO45-overexpressing mice, followed by tissue collection and pathology assessment via quantitative real-time PCR, histology, and immunohistochemistry.