Here, we have reported a case-control study investigating the association between XRCC1 codons Arg194Trp, Arg280His, and Arg399Gln and the development of NSCLC. NSCLC patients (N = 245) and healthy controls (N = 257) were randomly selected from the Huaihe Hospital between March 2012 and August 2014. DNA extracted from the patient and control blood samples were subjected to polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) to assess the genotyping of XRCC1 Arg194Trp, Arg280His, and Arg399Gln. Multivariate logistic regression analyses revealed an association between the expression of the AA genotype and A allele genotypes and a significantly increased risk of NSCLC, compared to the GG genotype [95% confidence interval (CI); Odd's ratio (OR) = 2.82 (1.141-5.86) and 1.67 (1.17-2.37), respectively]. The potential association between the A allele of XRCC1 Arg399Gln and the risk of NSCLC was more evident in smokers (95%CI; OR = 1.70; 1.11- 2.63). In conclusion, the XRCC1 Arg399Gln polymorphism was found to be associated with increased risk of NSCLC, especially in tobacco smokers.