Epstein—Barr virus (EBV) infection in tumor cells is generally restricted to the latent forms of viral infection. Switching the latent form of viral infection into the lytic form may induce tumor cell death. An important nuclear factor, nuclear factor (NF)-κB, is thought to play an essential role in EBV lytic infection; high levels of NF-κB can inhibit EBV lytic replication. In this study, we tested the effect of inducing EBV lytic replication using two NF-κB inhibitors: Bay11-7082 and Z-LLF-CHO, to reveal the possibility of targeting EBV-positive cancer therapy with these two NF-κB inhibitors. Our results showed that Bay11-7082 and Z-LLF-CHO reactivated EBV in a dose-dependent manner, thus resulting in EBV-positive 5-8F cell death. In contrast, there was no significant effect on EBV-negative HNE3 cells. When ganciclovir was used in combination with either Bay11-7082 or Z-LLF-CHO to treat 5-8F cells, the cytotoxic effect of the NF-κB inhibitor was amplified. The finding indicates that inhibiting the NF-κB activity of EBV-positive cells can induce lytic replication of EBV and cause lytic cytotoxicity against these cells.