Crosstalk of autophagy and ROS in multiple myeloma cells stimulated with doxorubicin / 中国病理生理杂志

ABSTRACT

AIM:To investigate the relationship of autophagy and reactive oxygen species ( ROS) in multiple myeloma cell line RPMI-8226 stimulated with doxorubicin.METHODS:The RPMI-8226 cells were stimulated with doxo-rubicin at different doses, and untreated cells were used as control.The protein expression of beclin 1 and LC3 was detec-ted by Western blot.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 cy-tometry, respectively.After treated with or without antioxidants tempol and N-acetyl-L-cysteine ( NAC) , the expression of beclin 1 and LC3 in RPMI-8226 cells was determined by Western blot.RESULTS:The protein levels of beclin 1 and LC3Ⅱ/LC3Ⅰwere increased in the RPMI-8226 cells stimulated with doxorubicin compared with untreated group.The ROS production was increased in the RPMI-8226 cells stimulated with 2 mg/L doxorubicin compared with untreated group.After treated with 3-MA, the ROS production and apoptosis in the RPMI-8226 cells stimulated with doxorubicin were increased compared with doxorubicin group.After treated with antioxidant NAC or tempol, the expression of beclin 1 and LC3 II/I in the RPMI-8226 cells stimulated with doxorubicin was decreased compared with doxorubicin 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 autophagy in multiple myeloma cells.