Determination of the anti-glioblastoma effect of artesunate and its potential targets through RNA-seq data analysis

Abstract Background: Glioblastoma (GBM) is an invasive brain tumor that lacks effective treatment methods. This study examined the effects and molecular mechanisms of Artesunate (ART) in GBM using both in vivo and in vitro methods and RNA sequencing (RNA-seq). Methods and Results: The effects of ART were assessed in vitro using GBM cells and in vivo using tumor-bearing nude mice. ART significantly suppressed GBM cell proliferation, facilitated apoptosis, and induced excessive reactive oxygen species (ROS) generation. However, ROS scavengers reversed the growth inhibitory and apoptotic effects of ART on GBM cells. In the mouse model of GBM, ART effectively inhibited cancer development without inducing toxicity. RNA-seq of ART-treated and untreated cells revealed 389 differentially expressed genes (DEGs), including 145 upregulated and 244 downregulated DEGs. Furthermore, the DEGs were applied to Gene Ontology and Kyoto Encyclopedia of Gene and Genomes analysis. The downregulated DEGs were mostly related to tumor-related pathways, such as the cell cycle, cell molecule adhesion, and extracellular matrix-receptor interaction. In addition, analyses using the Molecular Complex Detection algorithm and protein-protein interaction network were performed on those DEGs, and HMMR, CDC20, CCNB1, and THBS1, which may play key roles in ART-treated GBM cells, were identified. These results were validated via quantitative PCR and western blot analysis. Conclusions: Overall, our results illustrated the anti-GBM effects of ART and the possible mechanisms involved.