CDC25C表达下调的黑色素瘤细胞线粒体应激反应和线粒体途径凋亡情况观察OACSTPCD

Objective To investigate the effects of cell division cycle 25C(CDC25C)down-regulation on the mito-chondrial stress response and mitochondrial pathway apoptosis in melanoma B16 cells.Methods B16 cells were cul-tured and randomly divided into the transfection group,transfection control group and blank control group.In the transfec-tion group,B16 cells were transfected with the lentiviral plasmid with low-expression of CDC25C.In the transfection con-trol group,B16 cells were transfected with the lentiviral empty plasmid,while in the blank control group,B16 cells were cultured normally.Rhod-2/AM and MitoSOX fluorescence probe were used to detect the concentration of mitochondrial Ca2+ and mitochondrial reactive oxygen species(ROS)in B16 cells.The mRNA and protein expression levels of mitochon-drial stress response-related molecules mitochondrial caseinolytic protease P(ClpP)and Lon protease 1(LONP1),as well as the mitochondrial pathway apoptosis-related molecules Caspase-9 and Caspase-3 were detected using RT-qPCR and Western blotting.Results Compared with the transfection control group and the blank control group,the concentration of mitochondrial Ca2+ and mitochondrial ROS in B16 cells increased,and the mRNA and protein expression levels of ClpP,LONP1,and Caspase-9,and Caspase-3 increased in the transfection group(P<0.05 or P<0.01).Conclusion Down-regulation of CDC25C in melanoma B16 cells activates the mitochondrial stress response and subsequently triggers mito-chondrial pathway apoptosis.