Objective: To investigate the effect of inhibiting SATB2 gene expression on proliferation, apoptosis, invasion, and migration of oral squamous cell carcinoma cells and its mechanism. Methods: LipofectaminTM 2000 was used as vector, and negative control siRNA (negative group) and SATB2 specific siRNA group (inhibitory group) were transfected into human oral squamous cell carcinoma cell line CAL-27, and a blank group was set up, CCK-8 method was used to detect the proliferation of cells after siRNA was transfected into CAL-27 cells for 4 days, Western blotting were used to detect E-cadherin, STAT3, p-STAT3, and cleaved caspase-3 protein expression after siRNA was transfected into CAL-27 cells for 48 hours. Clone formation assay, Transwell chamber and flow cytometry were used to detect the cell clone formation rate, cell invasion and migration ability, and cell apoptosis rate. Results: The expression of SATB2 protein in CAL-27 cells transfected with siRNA was significantly lower compared with the blank group (P<0.05). Compared with the control group, the proliferation, invasion and migration of the cells in the inhibitory group decreased significantly, the apoptosis rate increased significantly, the expression of E-cadherin and cleaved caspase-3 protein increased significantly, and the expression of p-STAT3 protein was decreased significantly (P<0.05). Conclusion: Inhibition of SATB2 gene expression by RNA interference may inhibit the growth of oral squamous cell carcinoma cells, and the mechanism is related to down-regulation of STAT3 signal.