Objective To investigate the effect of thymopeptide α1 combined with FOLFOX4 chemotherapy regimen in the treatment of elderly patients with advanced gastric cancer. Methods A total of 102 elderly patients with gastric cancer were selected as the study subjects and randomly divided into a conventional group or an observation group, with 51 cases in each group. The conventional group was treated with FOLFOX4 chemotherapy regimen, based on which the observation group was treated with thymopeptide α1 additionally, and both groups received 3 cycles of treatment. The total effective rate after treatment, serum cytokeratin 19 fragment antigen 21-1 (CYFRA 21-1), carcinoembryonic antigen (CEA), tissue polypeptide specific antigen (TPS), chemokine (C-C motif) ligand 5 (CCL5), tissue inhibitor of metalloproteinase (TIMP)-1, matrix metalloproteinase (MMP)-9, MMP-2, CD3+, and CD4+ levels and CD4+/CD8+ ratio before and after treatment, and the occurrence of toxic and side effects during treatment were compared between the two groups. Results After 3 cycles of treatment, in the observation group, the total effective rate of treatment was higher than that in the conventional group, the serum levels of CYFRA 21-1, CEA, TPS, CCL5, TIMP-1, MMP-9, and MMP-2 were lower than those in the conventional group, and the serum CD3+ and CD4+ levels and the CD4+/CD8+ ratio were higher than those in the conventional group (all P<0.05). The incidences of toxic and side effects during treatment, such as gastrointestinal reactions, abnormal liver function, neurotoxicity, and myelosuppression in the observation group were lower than those in the conventional group (all P<0.05). Conclusion Thymopeptide α1 combined with FOLFOX4 chemotherapy regimen has a definite curative effect on elderly patients with gastric cancer, which can effectively improve patients' immune function, reduce their tumor markers levels and tumor cell invasion ability, and lower the incidences of toxic and side effects.