SPP1 was found to be significantly upregulated in many kinds of malignant tumors, including gliomas. Considering that gene polymorphisms have been implicated in the development of gliomas, we performed an association study between SPP1 functional promoter region polymorphisms and glioma risk in a Chinese population. We found significant evidence of an association between SPP1 promoter polymorphisms and glioma risk. For the -155_156insG variant, the -155_156GG allele was found to be significantly associated with an increased risk of glioma (P=0.020, odds ratio (OR)=1.202, 95% confidence interval (CI): 1.028-1.408). Individuals with the genotype containing the GG allele had a 1.372-fold increased risk (P=0.006, OR=1.372, 95% CI: 1.095-1.719). Further stratified analyses suggested that a significant association existed in adult glioma patients, male subjects and in cases without a family history of cancer. Alternatively, the study of single-nucleotide polymorphism -443C/T in a recessive model revealed that the genotype CC+CT significantly decreased the risk of glioma when compared with TT (P=0.023, OR=0.774, 95% CI: 0.621-0.966). After the analysis of haplotypes, the haplotype -155_156GG/-443T was represented at a significantly higher frequency in cases (P=0.029, OR=1.192, 95% CI: 1.018-1.395). Cellular assay indicated that the transcriptional activity of the SPP1 promoter containing the -155_156GG allele significantly increased in glioma cells. Thus, variants of the SPP1 promoter might influence the risk of glioma by regulating promoter activity. Further analyses are necessary to validate our observation in larger samples or in other ethnic groups.