Abstract: Objective: To explain the role of c-Jun N-terminal kinase (JNK) in the apoptosis induced by apoptin gene in human Hela cell. Methods: The Hela cell was transiently transfected by pcDNAA3 plasmids which contained apoptin gene, and apoptosis was measured by flow-cytometry. JNK was detected using western blotting analysis at various time periods. Results: The apoptin gene could induce apoptosis of Hela cell in vitro at 48h after transfection. The level of phosphorylated form of JNK was increased at 24h and reached peak level at 48h after transfected with pcDNAA3. The mRNA of apoptin gene could be detected by RT-PCR at 24h after transfection. Survival cells were associated with time after transfection. Conclusion: Apoptin gene can induce apoptosis in Hela cell, and JNK can play an important role in the apoptosis induced by apoptin gene in vitro.