Objective Multiple biochemical and genetic strategies were used to downregulate early response gene NF-κB, whose activation controls squamous cell cancer-associated pathways. Study design NA cells, an oral cavity squamous cell cancer with high NF-κB activity, were cultured with biochemical NF-κB inhibitors TPCK and Calpain I inhibitor, as well as specific NF-κB antisense oligonucleotides. Cell proliferation was measured, as was NF-κB downregulation using functional luciferase reporter genes and electromobility shift assays. Results Significant downregulation of cell proliferation and NF-κB functional activity were demonstrated with either biochemical inhibitor, as well as the antisense oligonucleotides; however, additional nonspecific toxicities were observed with control antisense oligonucleotides. Conclusion and significance NF-κB is a potential target for squamous cancer treatment, as it is constitutively upregulated in vitro. Biochemical inhibition of NF-κB may be a viable treatment strategy for head and neck squamous cancers.