Non-Small Cell Lung Cancer (NSCLC) accounts for approximately 85% of bronchial cancers. Mutations affecting genes such as KRAS, NRAS, BRAF, and PIK3CA are associated with tumor histology and patient smoking history. This study, the first of its kind conducted in western Algeria, aimed to investigate these mutations in 64 NSCLC patients. It also contributed to the establishment of a tumor and somatic DNA bank to support cancer research. Of the 64 samples collected, 50 were suitable for molecular analysis, while 14 were excluded due to low DNA concentrations (<0.05 ng/µl). Among the 50 samples, 30% showed KRAS mutations: 24% had the G12D type (linked to smoking), 2% G12C, 2% Q61K, and 2% A146T. A statistically significant association was found between KRAS mutations and smoking status (P = 0.02014). Additionally, 20% of the patients had PIK3CA mutations: 3 had E542G, 2 E542K, and 5 E545K. Notably, five patients exhibited concurrent KRAS and PIK3CA mutations, a finding supported by previous studies. No BRAF V600E mutations were detected. Interestingly, the mutation frequency of PIK3CA in this Algerian cohort appears higher than what has been reported in international studies.