AIM: To investigate whether the antitumor component I of Agkistrodon venom can inhibit the proliferation and migration of human oral squamous cell carcinoma SCC4 and Cal27 cells and whether it inhibits their proliferation and migration through the ERK1/2/MAPK signaling pathway. METHODS: After different concentrations of AAVC-I (0, 2.5, 5, 10, 20, 40 μg/mL) were applied to human oral squamous cell carcinoma SCC4 and Cal27 cells for 24 hours, the cell proliferation ability was detected by CCK-8 and EDUC-488 experiments. Cell apoptosis was detected by flow cytometry, cell invasion ability was detected by Transwell assay, cell migration ability was detected by scratch assay, and expression of related proteins in MAPK signaling pathway was detected by Western blot assay. RESULTS: After different concentrations of AAVC-I (0, 2.5, 5, 10, 20, 40 μg/mL) were treated on human oral squamous cell carcinoma SCC4 and Cal27 cells for 24 hours, the experimental group was compared with the normal control group. With the increase of AAVC-I concentration, the proliferation, invasion and migration ability of SCC4 and Cal27 cells of human oral squamous cell carcinoma gradually weakened, and the percentage of apoptosis gradually increased. The expression levels of MEK1 and ERK1/2 in MAPK signaling pathway did not change significantly. However, phosphorylated MEK1 and phosphorylated ERK1/2 decreased with the increase of drug concentration. Its expression level was down-regulated.CONCLUSION: AAVC-I can inhibit the proliferation, invasion and migration of human oral squamous cell carcinoma SCC4 and Cal27 cells, and the mechanism of its inhibition may be closely related to ERK1/2/MAPK signaling pathway.