Objective The aim of this study was to evaluate the significance of ANXA7 gene promoter hypermethylation in Glioblastoma.Methods Sixty patients with Glioblastoma and 24 healthy donors were adopted in this study. DNA was isolated from cerebra tissue, and BS-PCR and MS-PCR methods were used to detect the status of ANXA7 gene methylation in health donors and newly diagnosed Glioblastoma patients in using PCR amplication and sequencing.Results The results indicated that the methylated rate of the ANXA7 gene in 5 healthy donors were 1.5%, 1%, 1%,1.5%,1% by BS-PCR and sequencing analysis. however, the methylated rate of the ANXA7 gene in 5 Glioblastoma patients were 92%,78.5%,86%,56%,90%. MS-PCR analysis showed the ANXA7 gene was unmethylated in cerebra tissue samples from health donors. Among 60 newly diagnosed Glioblastoma patients, 25 patients were found in ANXA7 gene methylation by MS-PCR, the positive rate was 41.67% (p=0.000). Furthermore, the positive rate of ANXA7 methylation in the group of high stage disease was higher than in the group of low stage.Conclusions The aberrant methylation of the ANXA7 gene was perhaps involved in the occurrence of glioblastoma.