Background <p>YTH domain-containing protein 1 (YTHDC1) has been confirmed to be involved in regulating esophageal cancer (EC) progression. However, whether YTHDC1 mediates the radioresistance of EC and its potential molecular mechanism still need to be further explored.</p> Methods <p>The mRNA and protein levels of YTHDC1, ubiquitin-specific peptidase 10 (USP10) and polo-like kinase 1 (PLK1) were determined by qRT-PCR and western blot. Cell proliferation, invasion, migration, radiosensitivity, and apoptosis were detected by CCK8 assay, transwell assay, colony formation assay, and flow cytometry. The level of autophagy-related marker LC3B was examined using western blot. The interaction between USP10 and YTHDC1 or PLK1 was assessed by RIP assay, Co-IP assay and ubiquitination assay. Animal experiments were performed to explore the role of YTHDC1 in vivo.</p> Results <p>YTHDC1 was increased expression in EC tissues and cells. Silencing of YTHDC1 suppressed EC cell proliferation, migration and invasion, while promoted apoptosis, radiosensitivity and autophagy. YTHDC1 could stabilize USP10 mRNA level, and USP10 increased PLK1 expression by deubiquitination. Further analysis showed that PLK1 overexpression reversed the regulation of YTHDC1 knockdown on EC cell proliferation, invasion, radiosensitivity and autophagy. Besides, YTHDC1 knockdown could inhibit EC tumorigenesis and improve radiosensitivity in vivo via inactivating USP10/PLK1 axis.</p> Conclusion <p>Targeted inhibition of YTHDC1/USP10/PLK1 axis may be an effective measure to inhibit EC progression and improve radiosensitivity.</p>