Apoptotic signal transduction of Jurkat cells by cross-linking anti-human DR5 monoclonal antibody

Objective To detect the effect of cross-linking anti-DR5 monoclonal antibody YM366EC on the proliferation of Jurkat cells, and to elucidate the mechanism of anti-tumor by anti-DR5. Methods Cell viability of Jurkat was assayed by MTT. Apoptosis of Jurkat cells induced by cross-linking YM366EC was determined with Annexin V/PI staining using flow cytometry. Bcl-2, Cyt-C, Caspase-8, Caspase-3, Bax and Caspase-9 were determined by Western blot analysis. Results Although inhibition of Jurkat cells were not fund by anti-DR5 monoclonal antibody alone, apoptosis of Jurkat cells were detected with cross-linking anti-DR5 antibody. Cross-linking anti-DR5 YM366EC antibody showed a decreased expression of Bcl-2 and an increased activation of Cyt-C, Bax, Caspase-8, Caspase-3, Caspase-9 by Western blot analysis. Conclusion Apoptosis of Jurkat cells can be induced by cross-linking anti-DR5 YM366EC antibody, and the apoptosis signal pathway is relevant to Cyt-C, Bax, Caspase-8, Caspase-3 and Caspase-9.