17-羟-岩大戟内酯B对人三阴性乳腺癌细胞增殖和凋亡的影响  

Study on the effects of 17-hydroxy-jolkinolide B on the proliferation and apoptosis of human triple-negative breast cancer cells

作　　者：龚飞 吴思明 徐磊 包亚男[1] 林宇[1] 潘思文 杨东兴 韩翠翠[1] GONG Fei;WU Siming;XU Lei;BAO Yanan;LIN Yu;PAN Siwen;YANG Dongxing;HAN Cuicui(College of Pharmacy,Qiqihar Medical University,Heilongjiang Qiqihar 161006,China;Dept.of VascularSurgery,the First Hospital of Qiqihar,Heilongjiang Qiqihar 161006,China)

机构地区：[1]齐齐哈尔医学院药学院,黑龙江齐齐哈尔161006 [2]齐齐哈尔市第一医院血管外科,黑龙江齐齐哈尔161006

出　　处：《中国药房》2023年第12期1415-1421,共7页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81903865);齐齐哈尔市科技计划联合引导项目(No.LHYD-202003);黑龙江省大学生创新创业训练计划项目(No.X202211230021)。

摘　　要：目的研究狼毒大戟活性成分17-羟-岩大戟内酯B(HJB)对人三阴性乳腺癌(TNBC)细胞MDA-MB-231、MDA-MB-468增殖及凋亡的影响。方法用0(空白对照)、5、10、20、40、80μmol/L的HJB分别处理MDA-MB-231、MDA-MB-468细胞24、48、72 h,采用MTT法检测各组细胞的增殖抑制率。以0(空白对照)、10、20、40μmol/L的HJB作用于上述2种细胞24 h后,采用激光共聚焦显微镜和流式细胞术检测细胞的凋亡情况和线粒体膜电位、活性氧水平的变化情况,并采用Western blot法检测B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)、细胞色素C(Cyt-C)、胱天蛋白酶3(caspase-3)、活化的caspase-3(cleaved caspase-3)、caspase-9、cleaved caspase-9蛋白的表达水平。结果与空白对照相比,5、10、20、40、80μmol/L的HJB均可显著升高MDA-MB-231、MDA-MB-468细胞的增殖抑制率(P<0.05),且有浓度和时间依赖趋势。10、20、40μmol/L的HJB作用24 h后,2种细胞凋亡增多,总凋亡率均显著升高(P<0.05);线粒体膜电位均显著降低(P<0.05);活性氧水平均显著升高(P<0.05);Bcl-2、caspase-3、caspase-9蛋白的表达均显著下调(P<0.05),Cyt-C、Bax、cleaved caspase-3、cleaved caspase-9蛋白的表达均显著上调(P<0.05)。结论HJB对MDA-MB-231、MDA-MB-468细胞的增殖具有抑制作用,并可诱导其凋亡。OBJECTIVE To study the effects of the active component 17-hydroxy-jolkinolide B(HJB)of Euphorbia fischeriana on the proliferation and apoptosis of human triple-negative breast cancers(TNBC)MDA-MB-231 and MDA-MB-468 cells.METHODS MTT assay was adopted to detect the inhibitory rate of MDA-MB-231 and MDA-MB-468 cells proliferation after treated with 0(blank control),5,10,20,40,80μmol/L HJB for 24,48 and 72 h.Laser confocal microscope and flow cytometry were adopted to detect the apoptosis,mitochondrial membrane potential(MMP)and reactive oxygen species(ROS)of above 2 kinds of cells after treated with 0(blank control),10,20,40μmol/L HJB for 24 h.Western blot assay was used to detect the expressions of B cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),cytochrome-C(Cyt-C),caspase-3,cleaved caspase-3,caspase-9 and cleaved caspase-9.RESULTS Compared with blank control group,5,10,20,40,80μmol/L HJB could significantly increase the inhibitory rate of MDA-MB-231 and MDA-MB-468 cells proliferation(P<0.05),in dose-and timedependent trend.After 24 h treatment of HJB(10,20,40μmol/L),the apoptosis of above 2 kinds of cells increased,and the total apoptotic rate increased significantly(P<0.05);the mitochondrial membrane potential decreased significantly(P<0.05);the level of ROS increased significantly(P<0.05);the protein expressions of Bcl-2,caspase-3 and caspase-9 were decreased significantly(P<0.05),while the protein expressions of Cyt-C,Bax,cleaved caspase-3 and cleaved caspase-9 were increased significantly(P<0.05).CONCLUSIONS HJB can inhibit the proliferation of MDA-MB-231 and MDA-MB-468 cells,and induce their apoptosis.

关 键 词：17-羟-岩大戟内酯B 三阴性乳腺癌 细胞凋亡 线粒体膜电位 活性氧 

分 类 号：R965[医药卫生—药理学]