Microarray analysis of gene expression differences in microglia after exposure to graphene quantum dots

Abstract

Graphene quantum dots (GQDs) have been broadly applied in biomedicine in recent years. So far, researches have reported that GQDs might contribute to the injury of the central nervous system (CNS), yet the latent toxicological mechanism is not clear. This study aims to investigate the underlying biological mechanism responsible for the neurotoxicity of nitrogen-doped GQDs (N-GQDs) and amino-functionalized GQDs (A-GQDs) by use of genome-wide transcription microarray. The findings showed that 174 and 1341 genes were altered significantly in the BV2 cells treated by 25 μg/mL N-GQDs and 100 μg/mL N-GQDs compared with the control, respectively. As for the BV2 cells exposed to 100 μg/mL A-GQDs, 1396 diversely expressed genes were detected. By comparing the 100 μg/mL N-GQDs exposed group with 100 μg/mL A-GQDs exposed group, the expression of 256 genes was extensively altered, including 58 upregulated genes and 198 downregulated genes. From Gene Ontology (GO) analysis, the altered genes were mainly enriched in functions of ion channel activation and cellular processes. Based on the KEGG pathway and signal-net analysis, the toxicity of GQDs in BV2 cells was closely related to calcium signaling pathway, cell cycle and endocytosis. And the pathways that the shared mRNAs involved all served as the crucial roles in the neurotoxicity of GQDs despite the chemical functionalization (N-GQDs or A-GQDs). In addition, the consequences from qRT-PCR, Western blot, intracellular calcium level measurements and comet assay further confirmed that calcium dyshomeostasis, DNA damage and cell cycle arrest were the key factors responsible for the GQDs-induced neurotoxicity through affecting several classical signaling pathways. In conclusion, our research will supply essential data for further studies on mechanisms of GQDs-induced neurotoxicity by use of genome-wide screening.