Objective: To investigate the core target genes of miR-29b-3p, and analyze the clinical significance of the core target genes in glioma. Methods: Bioinformatics analysis was used to predict and screen the target genes of miR-29b-3p. STRING and Cytoscape software were used to analyze the protein-protein interaction (PPI) of target genes. the differences expression and survival prognosis in glioma were analyzed by GEPIA and CGGA. Independent prognostic factors analyzed by univariate and multivariate Cox proportional hazards regression model. Results: 22 target genes of miR-29b-3p were predicted using LinkedOmics, miRDB, miRTarBase, TargetScan, and starbase databases. Through the construction of the PPI network, genes out of the network were removed, and a total of 16 genes were screened for further study of their clinical significance. Based on analysis of GEPIA and CGGA databases, COL2A1, DNMT3A, and DNMT3B were excluded. Through further analysis of the univariate and multivariate Cox proportional hazard regression model, finally identified three core target genes: SERPINH1, LOXL2, CDK6. Conclusion: Bioinformatics analysis showed that miR-29b-3p targeted three core genes such as SERPINH1, LOXL2, and CDK6 in glioma. The expression of these genes was different between brain normal tissues and gliomas, between different grades of tumor, IDH mutation status and 1p/19q codeletion status. Its high expression had adverse effects on overall survival and recurrence-free survival. These core target genes can be used as an independent prognostic factor.