Diagnostic value of apolipoprotein C-I (ApoC-I), transthyretin (TTR) and ApoC-III in gastric cancer were evaluated. Retrospective analysis methods were used to collect 60 patients with gastric cancer first diagnosed in The First Affiliated Hospital of Jiaxing University. There were 60 patients with chronic atrophic gastritis in the benign lesion group and 60 healthy individuals in the control group. The expression levels of serum ApoC-I, TTR and ApoC-III was detected by enzyme-linked immunosorbent assay. Differences existed in the expression levels of ApoC-I, TTR and ApoC-III in the gastric cancer group, benign lesion group and control group (P<0.001), with the expression levels of ApoC-I, TTR and ApoC-III in the gastric cancer group being lower than that of the benign lesion group (P<0.05), and the expression levels of ApoC-I, TTR and ApoC-III in the benign lesion group being lower than that of the control group (P<0.05). The expression levels of ApoC-I, TTR and ApoC-III in the gastric cancer group were to a certain degree correlated with the clinical stage, lymph node metastasis and differentiation of patients in the gastric cancer group (P<0.05). The specificity and negative predictive value of combined detection were proven to be higher than the separate detection of the three factors (P<0.05). The detection of serum ApoC-I, TTR and ApoC-III was of great significance in the diagnosis of gastric cancer and the estimation of its severity. The method of combined detection is worth a further in-depth study as it could improve the specificity of diagnosis and have a higher negative predictive value.