Background: The present study aim to investigate the mechanism of Qigesan (QGS) on migration and invasion of esophageal cancer cells. Methods: First, we detected the cells in the normal group and the QGS group using microarray, and analyzed the ontology function and signaling pathway of target genes, and screened out TGF-β as one of the important targets of QGS. The subsequent verification experiments were divided into four groups: control group, model group (TGF-β1 stimulation group), QGS group (TGF-β1+QGS group) and positive control group (TGF-β1+TGF-β1 receptor inhibitor group). The migration and invasion abilities, as well as the expressions of related proteins and mRNA of each experimental group were detected. The migration and invasion ability of cells were detected by cell scratch test, and the Gene expression levels of E-cadherin, Vimentin, Smad2 and Smad7 mRNA in TGF-β1/Smad pathway were detected by RT-qPCR. The expression of E-cadherin, Vimentin, p-Smad2/3, Smad2/3 and Smad7 were detected by WB. Results: The results showed that TE-1 cells were reversed from fibroblast induced by TGF-β1 to epithelial cells after being treated QGS with the concentration of 20ug/mL for 36 h. Besides, QGS inhibited the migration and invasion of TE-1 cells and down-regulated the Gene and protein expression of Vimentin, Smad2 mRNA and Vimentin, p-Smad2/3 and Smad2/3. Meanwhile, The gene and protein overexpression E-cadherin and Smad7 was observed in QGS-treated cells. Conclusions: Together, these data indicated that QGS interfered with the epithelial-mesenchymal transition (EMT) process of TE-1 cells and reduced the migration and invasion of TE-1 cells via regulating TGF-β pathway, which maybe provided idea for the treatment of esophageal cancer.