Cisplatin-based chemotherapy is the standard advanced esophageal squamous cell carcinoma (ESCC) treatment. However, the 5-fluorouracil plus L-leucovorin and oxaliplatin (FOLFOX) regimen is available in Japan as an alternative to cisplatin-based chemotherapy, but its efficacy and safety remain unclear. Hence, we aimed to evaluate patients with advanced ESCC who received FOLFOX therapy retrospectively. Patients with advanced ESCC who received FOLFOX therapy at 18 institutions between April 2019 and October 2020 were included. Fifty-two patients received first-line FOLFOX treatment, while 39 received later-line FOLFOX treatment. The median progression-free survival, median overall survival, and objective response rate were 3.8 months (95% confidence interval [CI]: 3.0-5.0), 13.9 months (95% CI: 10.3-17.9 months), and 35% in the first-line group and 2.4 months (95% CI: 2.1-3.8 months), 7.2 months (95% CI: 5.3-9.8 months), and 4% in the later-line group, respectively. The most frequent grade 3 or 4 adverse events were neutropenia (23%) and anemia (12%) in the first-line group and neutropenia (18%), anorexia (13%), anemia (10%), and nausea (10%) in the later-line group. First-line FOLFOX treatment is suitable for patients with advanced ESCC. However, FOLFOX has shown limited efficacy as a later-line treatment.