Abstract Background: Heat shock protein 70 (Hsp70) has been shown to act as a chaperone and be associated with a variety of tumors. We investigated HSP70-1 G+190C, HSP70-2 A+1267G, and HSP70-hom T+2437C polymorphisms to assess whether genetic variation in HSP70 plays a role in the occurrence and development of lung cancer. Methods: A case-control study was conducted using 159 patients with lung cancer and 202 control subjects. Genomic DNA was typed for HSP70 polymorphisms using polymerase chain reactions with restriction fragment length polymorphism (PCR-RFLP). Unconditional logistic regression was used to estimate the relative risks of lung cancer. Results: There were significant differences in genotype and allele distributions between patients and controls for the HSP70-1 G+190C polymorphisms with and without adjustment for age, gender, smoking history, drinking history and family history of cancer (p<0.05). No significant differences were found in the polymorphisms of HSP70-2 A+1267G and HSP70-hom T+2437C. The haplotype analysis showed that the G/A/C and C/G/T haplotypes were associated with a significantly increased risk of lung cancer compared to the G/G/T haplotype. After adjustments for other risk factors, such as age, gender, drinking history and family history of cancer, the interactions between the HSP70-1 and HSP70-hom genotypes and smoking were confirmed [I(AB), 2.56 and 5.12, respectively]. Conclusions: HSP70-1 G+190C may be a functional polymorphism and affect susceptibility to lung cancer, and homozygous C/C genotype may enhance the risk of lung cancer. In addition, smoking along with HSP70-1 G+190C and HSP70-hom T+2437C, may increase the risk of lung cancer. Clin Chem Lab Med 2010;48:1657–63.