The Notch signaling pathway is a mechanism that plays a role in the determination of cell fate during cell development. Signals between neighbor cells are amplified through the Notch receptors. Notch activity is related to general growth stages such as organogenesis and morphogenesis and has effects on cell differentiation, cell proliferation, and apoptosis. Lung cancer associated with degradation of proteins which regulate cellular activities such as cell growth, differentiation, proliferation and apoptosis or the loss of function of proteins due to mutations in the genes which that express these proteins. We aimed to determine the frequency of the Notch3 rs3815188 (C381T) and rs1043994 (G684A) polymorphisms and to investigate whether this gene is associated with genetic predisposition of development of lung cancer. In this study, DNA samples were extracted from the venous blood sample of 200 subjects (100 lung cancer patients and 100 controls). Notch3 rs3815188 (C381T) and rs1043994 (G684A) polymorphisms were determined using the restriction fragment length polymorphism method. A statistically significant difference was found between the patient and control groups for Notch 3 gene rs3815188 and rs1043994 polymorphisms when evaluated in terms of genotype (p = 0.002 and p < 0.001, respectively) and allele frequencies (p < 0.05). In conclusion, the rs3815188 variant and rs1043994 variant of the Notch3 gene is associated with lung cancer risk in patients of Turkish origin.