In order to explore the role of 14-3-3σ gene methylation in nasopharyngeal carcinoma pathogenesis, biopsy specimens of 75 nasopharyngeal carcinoma tissues and 25 normal nasopharyngeal mucosal tissues were harvested for research. Then, methylation specific PCR (MSP) experiment was introduced to reveal 14-3-3σ gene methylation status of tissues, and reverse transcriptional PCR (RT-PCR) assay was employed to quantify the expression of 14-3-3σ mRNA, further more, immunohistochemical staining was undertaken to assess the 14-3-3σ protein expression level. The MSP experiment results indicate 14-3-3σ gene methylation status in nasopharyngeal carcinoma tissues was 4 exhaustive methylation, 59 partial methylation and 12 unmethylation, while in normal nasopharyngeal mucosal tissues was 7 partial methylation and 18 unmethylation. The frequency of 14-3-3σ methylation in nasopharyngeal carcinoma tissues was significantly higher than that in normal nasopharyngeal mucosal tissues (84% vs 28%, χ2=28; P < 0.05). The results of RT-PCR and immunohistochemical staining discovered that 14-3-3σ gene exhaustive methylation leads to absent 14-3-3σ mRNA as well as protein expression in nasopharyngeal carcinoma tissues and normal nasopharyngeal mucosal tissues. Tissues characterized with partial methylation 14-3-3σ gene represented remarkable down-regulated 14-3-3σ mRNA and protein expression level when compared with tissues characterized with unmethylation 14-3-3σ gene. Graded correlation analysis discovered that 14-3-3σ gene hypermethylation status is positively correlated with NPC lymphoid node metastasis and NPC clinical staging. The research demonstrated that 14-3-3σ gene was frequently hypermethylated in nasopharyngeal carcinoma tissues and leads to decreased or depleted gene expression, 14-3-3σ expression level was correlated to nasopharyngeal carcinoma lymphoid node metastasis and NPC clinical staging.