〔Abstract〕 Objective To investigate the role mechanism of doublecortin(DCX) in glioma and screen DCX overexpression-related differentially expressed genes, so as to provide new targets for glioma targeted therapy. Methods Using rat glioma C6 cells as the target cells, LV-DCX-EGFP, LV-Cas9-Puro, and LV-Ctrl-EGFP lentiviruses were constructed. DCX overexpressing and control C6 cell lines were established through Cas9 dual-vector system transfection and screening. Transcriptome sequencing was used to obtain the differentially expressed gene profiles. iDEP and DESeq2 were applied to screen differentially expressed genes with the threshold of |Log_2FC|>2 and P0.7, indicating good diagnostic value. RT-qPCR validation showed that the mRNA expression levels of some upregulated genes(POU2F3, LPAR6, SREBF1) significantly increased ( P < 0. 001) , while the mRNA expression levels of some downregulated genes (UBB , ACTB , UBE2I) also significantly increased (P < 0. 001) , which might be relat⁃ed to transcriptional and post⁃transcriptional regulation. Conclusion Differentially expressed genes and their regu⁃latory networks related to DCX overexpression in glioma are successfully screened , providing a theoretical basis for revealing the role mechanism of DCX in glioma development and laying a foundation for the development of potential therapeutic strategies.