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Introduction The aim of this study was to assess the role of TNF-α 308 G > A gene polymorphism in children's overweight risk so as to correlate this polymorphism with anthropometric and biochemical variables. Materials and method A cross-sectional study was carried out on 188 Romanian children ages 5–18 years, who were classified into controls (Group 1; n = 109) and overweight children (Group 2; n = 79). Results Higher values of MUAC and TST (p < 0.001) were obtained in the overweight group. A significant association was found between TNF-α 308 G > A polymorphism and weight status in the studied population (p = 0.009). There was also a positive association between the variant genotypes (GA or AA) of TNF-α 308G>A gene polymorphism and weight status, which was more frequently found among normal weight than overweight children (74.5% versus 25.5%, respectively). The final logistic multivariable included five independent variables (TNF-α genotype, gender, cholesterol, ASAT, and ALAT), which were statistically significant predictors with negative/positive effects on children's overweight risk; this model explained 30% of the variance in the outcome variable. Conclusion The variant genotype of TNF-α 308G>A gene polymorphism was more frequent among normal weight children. In the presence of other covariates, such as age, gender, cholesterol, LDL cholesterol, ALAT, and glycemia, the TNF-α 308 G>A gene polymorphism remained an independent protective factor for children's overweight status. [ABSTRACT FROM AUTHOR]