Combining network pharmacology and RNA sequencing to reveal the mechanism of emodin for the treatment of human neuroblastoma.

Abstract

Neuroblastoma (NB), as a highly metastatic tumor, represents the most common pediatric extracranial malignancy. Emodin is a natural product extracted from several traditional Chinese medicines, exerting potent anti-cancer properties in various cancer types. However, the detailed mechanism of emodin in the treatment of NB remains unclear. Network pharmacology was employed to explore the mechanism of emodin for the treatment of NB. The cell proliferation markers, cell cycle, cell cycle-related genes, and DNA damage-relevant genes of SH-SY5Y cell were examined by a battery of assays. Animal xenografts were used to evaluate tumor inhibition effect of emodin and perform RNA sequencing. Binding affinity of emodin and essential signaling proteins was investigated using molecular docking and confirmed by western blot analysis. The expression of epithelial-mesenchymal transition markers was also examined by western blot. Network pharmacology uncovered that emodin regulated cell cycle, p53 pathway, and PI3K/AKT pathway in NB. Emodin suppressed the cell proliferation in vitro by inducing the S phase arrest in SH-SY5Y cells, and the animal xenografts confirmed the anti-cancer activity of emodin in vivo. Further RNA-sequencing investigation showed that PI3K/AKT signaling is a potential pathway with emodin treatment. Our results validated that this signaling was indeed suppressed in the emodin-mediated anti-NB process, and molecular docking demonstrated that emodin bound strongly to PI3K and AKT1. And, emodin inhibited the metastasis of SH-SY5Y cells in vitro. Emodin restrained tumor growth by inducing S phase arrest and inhibited metastasis through inhibiting the PI3K/AKT signaling in SH-SY5Y cells.