To explore the overall situation of terahertz wave inhibition of K562 cell viability gene expression and reveal its possible key signaling pathways. In this study, terahertz wave was used to intervene K562 cells, RNA sequencing technology was used to analyze the overall differences in gene expression changes, and bioinformatics method was used to analyze the enrichment and clustering characteristics of differentially expressed genes. Under certain conditions, 1131 differentially expressed genes were screened, among which 216 expressed genes were up-regulated and 915 expressed genes were down-regulated. The results of gene ontology enrichment analysis showed that differentially expressed genes were mainly concentrated in DNA-binding transcriptional activator activity, RNA polymerase-DNA binding, transcription factor activity, protein folding, covalent chromatin modification and histone modification, etc. The signaling pathway analysis results of Kyoto Encyclopedia of Genes and Genomes showed that the differentially expressed genes were mainly concentrated in mitogen-activated protein kinase, C-type lectin receptor, Notch, Janus-activated kinase singal transducers and activators of transcriprion and transforming growth factor-β signaling pathways which are closely related to the proliferation and apoptosis of K562 cells. The study has certain reference value for revealing the effect of terahertz wave on organism.