摘要
目的:探讨β-谷甾醇抑制耐阿霉素人类髓性白血病细胞K562/ADR增殖、促进细胞凋亡与分化的效果及其可能机制。方法:使用细胞毒性试验计算β-谷甾醇对K562/ADR细胞的半数抑制率(IC50),采用CCK-8法检测细胞增殖率,采用流式细胞术检测细胞凋亡率与线粒体膜电位下降率,使用分光光度法检测细胞上清中SOD活力与MDA含量,使用荧光试剂盒检测细胞ROS强度,使用联苯胺染色检测红细胞分化程度,使用蛋白质印迹(Western blot)检测珠蛋白转录因子1(GATA-1)、β-珠蛋白(β-globin)和核因子e2相关因子2(NF-E2)蛋白表达水平。结果:β-谷甾醇对K562/ADR细胞的半数抑制浓度的IC50为163.64μmol/L。与空白组相比,3个浓度的β-谷甾醇均能抑制细胞增殖、促进细胞凋亡与红细胞分化,降低SOD活力、线粒体膜电位,增加MDA与ROS含量,升高GATA-1、β-globin和NF-E2蛋白表达水平(P<0.01)。结论:β-谷甾醇能抑制K562/ADR细胞增殖,促进细胞凋亡,并且诱导红细胞分化,其机制可能是通过促进ROS积累引起氧化应激失衡。
Objective:To investigate the effect of β-sitosterol on proliferation,apoptosis and differentiation of CML cell line K562/ADR and its possible mechanism.Methods:Cytotoxicity test was used to calculate the half inhibition rate concentration(IC 50)of β-sitosterol on K562/ADR cells.CCK-8 method was used to detect the cell proliferation rate.Flow cytometry was used to detect the cell apoptosis rate and mitochondrial membrane potential decline rate.Spectrophotometry was used to detect the SOD activity and MDA content in cell supernatant.Fluorescence kit was used to detect the ROS intensity.The erythrocyte differentiation was detected by benzidine staining,and the protein expression levels of GATA-1,β-globin and NF-E2 were detected by Western blot.Results:The IC 50 of β-sitosterol on K562/ADR cells was 163.64μmol/L.Compared with control group,β-sitosterol at three concentrations inhibited cell proliferation,promoted cell apoptosis and erythrocyte differentiation,decreased SOD activity and mitochondrial membrane potential,increased MDA and ROS content,and increased GATA-1,β-globin and NF-E2 protein expression levels(P<0.01).Conclusion:β-sitosterol can inhibit the proliferation of K562/ADR cells,promote cell apoptosis,and induce erythrocyte differentiation.The mechanism may be that β-sitosterol causes oxidative stress imbalance by promoting ROS accumulation.
作者
赵冰洁
车虹
胡琏
易文静
侯著法
ZHAO Bingjie;CHE Hong;HU Lian;YI Wenjing;HOU Zhufa(Department of Hematology,Affiliated Hospital of Chengdu University of Traditional Chinese Medicine,Sichuan Chengdu 610072,China)
出处
《现代肿瘤医学》
CAS
2024年第4期641-645,共5页
Journal of Modern Oncology
关键词
慢性粒细胞白血病
Β-谷甾醇
红细胞分化
氧化应激
体外
chronic myelogenous leukemia
β-sitosterol
erythrocyte differentiation
oxidative stress
in vitro
