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  

Effects of down-regulation of CDC25C on mitochondrial stress response and mitochondrial pathway apoptosis in melanoma cells

作　　者：缪欣宇 栗彦飞 郑方园 刘海龙 张瑶尧 晁耐霞[1] 莫发荣[1,2] MIAO Xinyu;LI Yanfei;ZHENG Fangyuan;LIU Hailong;ZHANG Yaoyao;CHAO Naixia;MO Farong(School of Basic Medical Sciences,Guangxi Medical University,Nanning 530021,China;不详)

机构地区：[1]广西医科大学基础医学院,南宁530021 [2]广西高校人体发育与疾病研究重点实验室

出　　处：《山东医药》2024年第2期31-34,38,共5页Shandong Medical Journal

基　　金：广西自然科学基金项目(2023GXNSFAA026070,2018GXNSFAA281071)。

摘　　要：目的探讨细胞分裂周期蛋白25C(CDC25C)对黑色素瘤B16细胞线粒体应激反应和线粒体途径凋亡的影响。方法取黑色素瘤B16细胞进行培养,将细胞随机分为转染组、转染对照组、空白对照组。转染组转染CDC25C低表达慢病毒质粒,转染对照组转染慢病毒空质粒,空白对照组正常培养。利用Rhod-2/AM与MitoSOX荧光探针检测细胞线粒体内Ca^(2+)浓度及线粒体活性氧(ROS),RT-qPCR法和Western blotting法分别检测细胞线粒体应激反应相关分子ClpP、LONP1及线粒体途径凋亡相关分子Caspase-9、Caspase-3 mRNA和蛋白表达。结果与转染对照组和空白对照组相比,转染组细胞线粒体Ca^(2+)及ROS含量增加,ClpP、LONP1、Caspase-9、Caspase-3 mRNA及蛋白表达水平升高(P<0.05或<0.01)。结论CDC25C表达下调激活黑色素瘤B16细胞发生线粒体应激反应,并诱导线粒体途径的细胞凋亡。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 Ca^(2+)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 Ca^(2+)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.

关 键 词：黑色素瘤 细胞分裂周期蛋白25同源蛋白C 线粒体应激反应 线粒体钙离子超载 线粒体活性氧 细胞凋亡 

分 类 号：R739.5[医药卫生—肿瘤]