The Study Of Combination Of Arsenic Trioxide With All-trans Retinoic Acid On Induction Of NF-kb,ROS In NB4 Cells

Objective:To study the effects of combination of all-trans retinoic acid with arsenic trioxide on the cell growth , differentiation, apoptosis and the expression of NF-κB, ROS in NB4 cells , and to analysis mutual effects of all-trans retinoic acid and arsenic trioxide at the different concentrations. Methods:NB4 cells which grew in logarithmic phase were divided into three groups. The frist group were treated with lumol/L ATRA alone , 1μmol/L ATRA plus 0.5μmol/L As₂O₃ , 1μmol/L ATRA plus 2.0μmol/L As₂O₃ respectively. The second group were treated with 0.5μmol/L As₂O₃ alone , 2.0μmol/L As₂O₃ alone and in combination with 1μmol/L ATRA respectively . The third group were treated without agents.1) Methods of Trypan blue exclusion assay and cell growth curve were used to observe proliferation of NB4 cells which were harvested at zero hour, twenty-four hours, forty-eight hours and seventy-two hours respectively.2) Fluctuation of CD11b was detected by flow cytometry to test differentiation of NB4 cells which were harvested at four hours, twelve hours, twenty-four hours, forty-eight hours and seventy-two hours respectively.3) NB4 cells were harvested at twenty-four hours, forty-eight hours and seventy-two hours respectively. Cell apoptosis was observed by DNA laddering assay.4) NB4 cells which were harvested at 48 hours were cultured with P65 nuclear factor-κB (NF-κB) monoclonal antibody and secondary antibody. Fluctuation of NF-κB was detected by flow cytometry.