Abstract: The neurotoxic mechanism of PrP106-126 still remains uncertain. Herein PrP106-126-induced neurotoxicity in mouse neuroblastoma cell line N2a, for the first time we found that PrP106-126 induced p65 subunit of nuclear transcription factor NF-κB translocated into the nucleus by Western Blot assay and activated NF-κB signaling pathway. Pretreatment with nuclear translocation inhibitor NF-κB SN50 attenuated PrP106-126-induced NF-κB activity, using apoptotic DNA Ladder assay and flow cytometry (FCM) assay with Annexin V-FITC/PI double-staining, we also found that attenuated NF-κB activity was accompanied by reduced apoptotic effect by PrP106-126. The results showed that NF-κB signaling pathway was involved in PrP106-126-induced neurotoxicity, and NF-κB activation showed pro-apoptotic effect in N2a cells. The in vitro study on PrP106-126-activated NF-κB signaling pathway and its pro-apoptotic effect on N2a cells were significantly important to elucidate the neurotoxic mechanism of prion.