Untargeted UPLC-MS-based metabolomics analysis reveals the metabolic profile of intrahepatic cholangiocarcinoma process and the intervention effect of Osthole in mice

Abstract

Intrahepatic cholangiocarcinoma (ICC) is the second most common malignant liver tumor after hepatocellular carcinoma (HCC). It poses a significant risk to human health as it is highly malignant, insidious, and prone to recurrence after surgery. However, current diagnostic techniques and treatments for ICC are limited. Therefore, it is critical to investigate biomarkers of ICC for the discovery of novel drugs that can effectively treat ICC. In a previous study on the molecular mechanisms of ICC, proto-oncogenes AKT and Yap were found to often be overexpressed or activated in ICC, and the co-expression of AKT/YapS127A could promote the formation of ICC in mice. In this study, co-expression of AKT/YapS127A in an ICC mouse model was established via hydrodynamic transfection. To identify ICC biomarkers, we investigated the inhibitory effect of Osthole on the formation of ICC in mice induced with the co-expression of AKT/YapS127A. Furthermore, we preliminarily established the pathogenesis of ICC and the mechanism of Osthole. The metabolomic profiles for different stages in ICC formation process and Osthole treatment were examined via UHPLC-Q-TOF-MS/MS. The morphology (hematoxylin and eosin and Sirius red staining) and immunohistochemistry (CK19, Ki67, Vimentin, and α-SMA) results demonstrated that the ICC mouse model was useful. The ICC formation process was divided into three stages: early (1–2 weeks), formative (3–4 weeks), and late (5–7 weeks) stages. For the metabolomic profiles of ICC formation process, 11 differential metabolite species were identified at the early stage, 14 at the formative stage, and 20 at the late stage. The model group mice mainly showed dysregulation of amino acids and lipid metabolism compared with the control group mice. For the metabolomic profiles of Osthole treatment, 14 potential biomarkers with reduced levels were preliminarily identified, which were also linked to the dysregulation of amino acids and lipid metabolism. Combined with the results of Western Blotting experiments, it was found that Osthole can stop tumors from progressing by altering the AKT/mTOR signaling pathway with changes in amino acid and lipid metabolism. This study can provide a potentially valuable metabolic basis for the early detection, treatment, and prognosis of ICC, as well as the clinical application of Osthole in ICC.

Keywords

• Intrahepatic cholangiocarcinoma
• AKT/YapS127A co-expression
• Osthole
• Metabonomics
• UHPLC-Q-TOF-MS/MS