Crosstalk Of Autophagy And ROS In Multiple Myeloma Cells Stimulated Wit Doxorubicin

AIM: To investigate the relationship of autophagy and reactive oxygen species(ROS) in multiple myeloma cell(MM) line RPMI-8226 stimulated with doxorubicin.METHODS: The RPMI-8226 cells were stimulated by doxorubicin with different dose, untreated cells were used as control. The protein expression of beclin1 and LC3 was detected by Western blotting. ROS production was analyzed by DCFH-DA fluorescence staining. After treated with or without 3-methyladenine(3-MA), the ROS production and apoptosis in RPMI-8226 cells were determined by DCFH-DA and flow cytometry, respectively. After treated with or without antioxidant Tempol and N-acetyl-L- cysteine(NAC), the expression of beclin1 and LC3 in RPMI-8226 cells was measured by Western blotting.RESULTS: The protein levels of beclin1 and LC3B/LC3 A were increased in RPMI-8226 cells stimulated with doxorubicin compared with untreated group. When the concentration of doxorubicin at 2 mg/L induced, the ROS production was increased in RPMI-8226 stimulated withdoxorubicin compared with untreated group. After treated with or without3-MA,The ROS production in RPMI-8226 cells stimulated with doxorubicin was increased compared with control group. After treated with or without antioxidant NAC or Tempol, The expression of beclin1 and LC3II/I in RPMI-8226 cells stimulated with doxorubicin was decreased compared with control group.CONCLUSION: The autophagy and ROS levels are increased in RPMI-8226 cells stimulated with doxorubicin. Inhibition of autophagy increases the ROS production and apoptosis of RPMI-8226 cells stimulated with doxorubicin. Inhibition of ROS production reduces doxorubicin induced by autophagy expression of multiple myeloma cells.