The tumor suppressor p53 plays a crucial role in maintaining genomic stability and tumor prevention. Murine double-minute 2 (MDM2) oncoprotein plays a pivotal role in regulating p53, and the single-nucleotide polymorphism (SNP) 309T/G SNP in the promoter region of Mdm2 has been shown to be associated with increased risk of cancer. We investigated the association between Mdm2-309 promoter polymorphism, plasma MDM2 levels, and risk of laryngeal squamous cell carcinoma (LSCC). In this case-control study, 146 patients with LSCC, 61 patients with vocal leukoplakia, and 212 healthy controls were genotyped for the Mdm2-309 T/G gene using pyrosequencing. Plasma MDM2 levels were also analyzed by enzyme-linked immunosorbent assay (ELISA). Patients with LSCC had a significantly lower frequency of GT at Mdm2-309 (odds ratio [OR]=0.50, p=0.02) than controls. The proportion of GT heterozygotes in advanced stage cases were less than that in the initial stage patients (OR: 0.36 vs. 0.63; p=0.007 and 0.16). The same result was found between cases with and without lymph node metastases (OR: 0.45 vs. 0.52; p=0.075 and 0.04). Moreover, the plasma Mdm2 concentrations of LSCC patients (343.36±14.8 pg/mL) were significantly higher than those in controls (255.76±8.2 pg/mL; p<0.01) and vocal leukoplakia patients (301.42±8.6 pg/mL; p<0.05). Patients in advanced stages and with lymph node metastasis had higher plasma MDM2 levels, while the GT genotypes (308.06±18.9 pg/mL; p=0.037) had lower MDM2 plasma levels than the TT genotypes (369.00±25.2 pg/mL). The Mdm2 SNP309 G allele is implicated as an important LSCC and a vocal leukoplakia protective factor in the Chinese Han Population, and the proportion of GT genotype was lower in advanced LSCC patients and lymph node metastasis patients. Moreover, Mdm2-309 GT genotype patients had a lower plasma MDM2 level than the TT genotypes.