NF-κB has been recognized as one of the factors responsible for the development of cancer; however, the mechanism by which high expression of NF-κB contributes to the progression of human oesophageal squamous cell cancer (ESCC) is not fully understood. In our investigations, NF-κBP65 was overexpressed in human ESCC tissues, especially in ESCC tissues with deep invasion and lymph node metastasis. Suppression of NF-κBP65 by siRNA decreased the invasion and proliferation ability of EC9706 cells in vitro. Furthermore, siRNA-mediated NF-κBP65 knock-down could lead to the downregulation of MMP-9, a metastasis-related gene. Reduced E-cadherin is a hallmark of invasive carcinomas that have acquired epithelial–mesenchymal transition (EMT) phenotypes and Vimentin is another molecule that is used widely as a marker of the EMT. We found upregulation of E-cadherin expression and downregulation of Vimentin was induced by NF-κBP65 siRNA, which suggests that NF-κBP65 siRNA could inhibit the invasion and proliferation ability of ECSS through attenuating the expression of MMP-9 and EMT. Thus, ESCC NF-κBP65 could be a useful target for cancer prevention and therapy.