<p>This study investigated the effects and mechanisms of action of dexmedetomidine (DEX) on the proliferation, migration, and invasion of esophageal squamous cell carcinoma (ESCC) cells. Network pharmacology and bioinformatics analyses identified core targets, including vascular cell adhesion molecule-1 (<i>VCAM-1</i>), chemokine receptor 2 (<i>CXCR2</i>), matrix metalloproteinases (<i>MMPs</i>) 1, 2, and 3, and coagulation factor II (thrombin) receptor (<i>F2R</i>). Among these genes, <i>F2R</i> was associated with poor prognosis in these patients. The experimental validation was performed using the TE13 human esophageal cell line, which were exposed to different concentrations of DEX (Experiment 1) and transfected with a lentivirus to generate a model of coagulation <i>F2R</i> overexpression (Experiment 2). Through a combination of techniques, including clone formation, scratching, MTT, Transwell assays, western blotting, as well as quantitative real-time PCR (qRT-PCR). Experiment 1 demonstrated DEX could significantly decreased TE13 cell proliferation, migration, and invasion abilities. Moreover, the levels of MMP1, MMP2, F2R, VCAM-1, and MMP3 proteins were significantly reduced after DEX treatment, whereas CXCR2 levels were significantly increase. Experiment 2 selected <i>F2R</i> as the target of DEX for mechanistic studies. DEX can reverse the malignant progression of TE13 cells overexpressing F2R. Western blotting revealed a significant decrease in the expression of F2R/ extracellular signal-regulated kinase (ERK) ERK1/2-related pathway proteins after DEX treatment (<i>P</i> &lt; 0.05). The foregoing results underscore DEX can inhibit the proliferation, migration, and invasion of ESCC cells, and its mechanism of action may be related to inhibition of the F2R/ERK1/2 pathway.</p>