〔Abstract〕 Objective To study the effect and mechanism of transcription factor SPIBon proliferation, migration and invasion of esophageal cancer cells after regulating the expression of SPIB protein by gene expression regulation technology. Methods The expression levels of SPIB mRNA and protein in esophageal cancer cell lines were detected by Western blot and qRT-PCR, respectively. SPIB expression was down-regulated by siRNA in KYSE150 cells, and the proliferation and cell cycle progression were detected by Cell Counting Kit-8 and flow cytometry, respectively. The KYSE30 cell lines with stable overexpression of SPIB were established and the effects of SPIB on cell proliferation and clone formation were detected. The effects of SPIB on the migration and invasion of esophageal cancer cells were detected by cell scratch and Transwell system, and the expression levels of N-cadherin and E-cadherin proteins were detected by Western blot. Results SPIB was highly expressed in esophageal cancer cell lines. Down-regulation of SPIB could inhibit cell proliferation and cell cycle progression, and decrease the expression of Cyclin D1 proteins. Overexpression of SPIB enhanced the proliferation and clone formation of esophageal cancer cells. Knockdown of SPIB protein expression weakened the migration and invasion ability of esophageal cancer cells, and reduced the expression of N-cadherin protein. Conclusion SPIB, as a pro-cancer factor, can promote the proliferation, migration and invasion of esophageal cancer cells.