| Liver cancer is a malignant disease that causes hundreds of thousands of deaths every year.Metastasis and recurrence of liver cancer have always been the difficulties in treatment.Therefore,it is urgent to develop anti-liver cancer drugs with better curative effect and lower toxic and side effects.Nur77 is an early gene product encoded by NR4A1 and belongs to the orphan nuclear receptor.It has been reported that Nur77 is significantly up-regulated in a variety of tumor tissues,such as pancreatic cancer,liver cancer and colon cancer,indicating that Nur77 has a pro-survival function in these tumors.Many antitumor drugs in clinical use contain indole skeleton in their chemical structure,which has attracted much attention due to its strong pharmacological activity and high modification degree.However,no anti-liver cancer indole compounds based on Nur77 have been developed so far.Therefore,this paper aims to screen out anti-liver cancer indole compounds targeting the nuclear receptor Nur77 with strong biological activity and low toxicity.First,we screened two indole compounds which named as z-e and z-i from a batch of candidate compounds by MTT method,which had good cytotoxic effects on liver cancer cells but had little effect on normal liver cells.Then,using the computer simulation of molecular docking technology and in vitro purification of Nur77 ligand binding domain(Nur77-LBD)protein,the molecular interaction analysis experiments confirmed that compounds z-e,z-i and Nur77-LBD interbind.Then,clonal formation and cell scratch experiments showed that the two compounds could inhibit the proliferation and migration of HCC cells.Further cell morphological analysis and Annexin V-FITC/PI assay showed that the compound could induce apoptosis,and the effect of z-e was better than that of z-i.Finally,we found that the compound degraded Nur77 through the proteasomal pathway and also inhibited cell cycle-related proteins and activated endoplasmic reticulum stress-related proteins.In this study,we also found that the regulation of cell cycle and endoplasmic reticulum stress-related proteins by the compound is dependent on the nuclear receptor Nur77. |
