Ciprofol exerts anti-tumour effects in hepatocellular carcinoma through the Raf-MEK-ERK signalling pathway.

Abstract

Background: Hepatocellular carcinoma (HCC) is one of the most prevalent malignant tumours of the digestive system and the third leading cause of cancer-related deaths worldwide. As the most common type of primary liver cancer, HCC is associated with poor prognosis despite advancements in treatment options such as radical resection, liver transplantation, and adjuvant therapies. Surgical resection remains the cornerstone of HCC treatment; however, postoperative recurrence and metastasis pose significant challenges to patient survival. Intraoperative factors, including immune suppression and the use of certain anaesthetics, have been implicated in tumour cell dissemination and recurrence. While anaesthetic agents like propofol are known to influence tumour cell proliferation, differentiation, and apoptosis. Ciprofol, a novel intravenous anaesthetic, has demonstrated clinical safety and efficacy, but its potential impact on HCC progression and underlying mechanisms requires further exploration. This study aims to explore how ciprofol affects the behaviour of HCC cells and the underlying mechanisms.

Methods: Hep3B and HCCLM3 HCC cell lines were treated with varying concentrations of ciprofol. The cell numbers were measured at different time points using the Cell Counting Kit-8 (CCK-8) to find the active concentration. Proliferation was assessed by colony formation and 5-ethynl-2'-deoxyuridine (EdU) assays, whereas invasion and migration were tested using Transwell and wound healing assays. Subcutaneous xenograft and orthotopic liver transplantation models were used to study tumour growth in vivo, and a lung metastasis model was created to examine its effects on metastasis. RNA sequencing (RNA-seq) identified transcriptional changes after ciprofol treatment, and western blot and immunofluorescence (IF) validated these findings.

Results: Ciprofol inhibited the proliferation, migration, and invasion of Hep3B and HCCLM3 cells in a manner dependent on both time and dosage. It also reduced tumour growth and lung metastasis in mice. RNA-seq showed that ciprofol affected the MAPK/ERK pathway, which was confirmed by the reduced phosphorylation levels of Raf, MEK, and ERK, without affecting total protein levels.

Conclusions: Ciprofol inhibited the MAPK/ERK pathway by reducing the phosphorylation of Raf, MEK, and ERK, which may explain its inhibitory effects on HCC. The results of this study could guide the use of anaesthetic drugs in HCC surgery.