Objective To explore the methylation status of ppENK promoter region in glioma and discuss the abnormal ppENK gene promoter methylation gliomas occurrence and development, the relationship between role and clinical pathological material. Methods Methylation status of the ppENK promoter region was assayed in 32 glioma tissues and 10 normal brain tissues by nested polymerase chain reaction (nPCR) and Bisulfite sequence polymerase chain reaction (BSP).Results BSP showed that the ppENK promoter region was hypermethylated in 13 of the 32 glioma tissues and no methylation of ppENK promoter was detected in the 10 normal brain tissues. The promoter hypermethylation ratio of ppENK gene in glioma tissues (40.6%) was significantly higher than that in normal brain tissues (0%, P=0.015). Moreover, we found that the aberrant promoter methylation of ppENK gene was significantly correlated with tumor grade (P=0.006), but not correlated with age, sex and tumor types of patients (P=0.154, 0.961 and 0.694, respectively). The rate (21.1%) of ppENK promoter methylation in grades Ⅰ~Ⅱ gliomas was significantly lower than that (69.2%) in grades Ⅲ~Ⅳ gliomas (P<0.05).Conclusions ppENK promoter (CpG island) hypermethylation exists in glioma, which may correlates with the occurrence of glioma. The expression and promoter region methylation of ppENK gene in the gliomas are related to pathological grades, not related to the patient's age, gender and tumor size factors.