Pancreatic cancer (PC) is one of the most fatal malignancies, which has attracted scientists to investigate its etiology and pathogenesis. Nevertheless, the association between erythrocyte fatty acids and PC risk remains unclear. This study aimed to evaluate the association between levels of erythrocyte fatty acids and PC risk. The erythrocyte fatty acid compositions of 105 PC patients and 120 controls were determined by gas chromatography. Cases and controls were frequency matched by age and sex. Multivariable conditional logistic regression model and restricted cubic spline were applied to estimate the odds ratio with 95% confidence interval (OR, 95% CI) of erythrocyte fatty acids and PC risk. Our main findings indicated a significant negative association between levels of erythrocyte total monounsaturated fatty acids (MUFA) and n‐3 polyunsaturated fatty acids (n‐3 PUFA) and the risk of PC (ORT3‐T1 = 0.30 [0.14, 0.63] and ORT3‐T1 = 0.15 [0.06, 0.33], respectively). In contrast, erythrocyte n‐6 polyunsaturated fatty acids, specifically linoleic acid (LA) and arachidonic acid (AA) levels, were positively associated with PC incidence (RT1‐T3 = 4.24 [1.97, 9.46] and ORT1‐T3 = 4.53 [2.09, 10.20]). Total saturated fatty acid (SFA), especially high levels of palmitic acid (16:0), was positively associated with the risk of PC (ORT3‐T1 = 3.25 [1.53, 7.08]). Our findings suggest that levels of different types of fatty acids in erythrocytes may significantly alter PC susceptibility. Protective factors against PC include unsaturated fatty acids such as n‐3 PUFA and MUFA.