Objective To investigate the effect of VRK1 expression downregulated by siRNA on BANF1 expression and the proliferation and migration abilities of esophageal squamous cell carcinoma (ESCC) cells. Methods VRK1 siRNA was transfected into EC109 and EC1 cell lines to interfere VRK1 expression. Western blot was used to detect the interference efficiency of VRK1 siRNA and BANF1 expression in esophageal carcinoma cells after siRNA interference. CCK-8 and flow cytometry were performed to detect the effect of VRK1 expression on cell proliferation. Transwell assay was used to observe the changes of cell migration ability. Results After VRK1 siRNA transfection, VRK1 expression were down-regulated for 62.40% and 52.14%, and BANF1 protein expression were down-regulated significantly for 24.51% and 52.87% in EC109 and EC1 cell lines, respectively. The proliferation of EC109 and EC1 cell lines were decreased 12h after transfection, with the highest inhibition rate at 36h(19.41% and 20.10%). The cells percentages in G2 and G1 phases were decreased while that in S phase was increased after down-regulation of VRK1 and BANF1 expression. The number of cell migration was decreased in the experimental group, compared with control group (P < 0.01). Conclusion Down-regulation of VRK1 expression by siRNA could inhibit the proliferation and migration abilities of esophageal squamous carcinoma cells, which may be achieved by down-regulating BANF1 expression.