Background: Gastric adenocarcinoma is the most common type of gastric cancer all around the world. The epithelial cells of stomach tissue are influenced by environmental factors and genetic disorders. P53 is the most remarkable gene that controls the growth of cells. Mutation in some nucleotides of the P53 gene increases the genetic instability and is assumed as an important prognostic factor in gastric cancer. More than 90% of mutations occur in the exons 5-8 of p53. Objectives: This study was conducted in Kohgiluyeh and Boyer Ahmad (K&B) province (Southwest Iran) to determine the rate of R282W P53 gene mutation of exon 8 in gastric cancer. Methods: This case-control study was conducted on 90 subjects that were divided into two groups (each including 45 patients and 45 controls). The samples were randomly collected from the tissue bank of the pathology laboratory in Yasuj city and then were transferred to the Cellular and Molecular Research Center. DNA extraction was performed by the DNA extraction kit. Molecular analysis on exon 8 was performed by the PCR-RFLP method and using the MspI restricting enzyme. Data were analyzed by descriptive statistics and bivariate correlation tests. Results: No difference was found between the two groups concerning age, gender, and education level. The prevalence of R282W P53 gene mutation on exon 8 in the cancer group was 17.8% (8/45), while no mutation was found in the control group. Conclusions: According to the results, the R282W P53 gene mutation on exon 8 may play an important role in the development of gastric cancer in Yasuj district, Southwest Iran.