^(125)I粒子联合AZD1152对三阴性乳腺癌细胞增殖和凋亡的影响  

The combination of^(125)I particles and AZD1152 efficiently inhibited proliferation and promoted apoptosis of a triple negative breast cancer cell line

作　　者：张月[1] 王耀一[1] 武雪亮[1] 张志生[1] 杨修明 姜洋 乔志飞[1] 梁晚平[1] 薛军[1] ZHANG Yue;WANG Yaoyi;WU Xueliang;ZHANG Zhisheng;YANG Xiuming;JIANG Yang;QIAO Zhifei;LIANG Wanping;XUE Jun(Department of Mammography Surgery,the First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,Hebei,China)

机构地区：[1]河北北方学院附属第一医院乳腺外科,河北张家口075000

出　　处：《癌变．畸变．突变》2022年第4期295-299,306,共6页Carcinogenesis,Teratogenesis & Mutagenesis

基　　金：张家口市2018年度科学研究计划项目(1821065D,1821066D)。

摘　　要：目的:研究^(125)I粒子联合Aurora激酶抑制剂AZD1152对三阴性乳腺癌(TNBC)细胞MDA-MB-231增殖和凋亡的影响。方法:试验设对照组(常规培养)、^(125)I粒子照射组(照射组)、1μmol/L AZD1152作用MDA-MB-231细胞48 h组(抑制组)、^(125)I粒子照射与1μmol/L AZD1152联合作用MDA-MB-231细胞48 h组(联合组)。采用免疫荧光检测细胞多核形成;四甲基噻唑蓝(MTT)法检测细胞增殖抑制率;CCK-8检测细胞活力;流式细胞术PI单染检测细胞周期;流式细胞术Annexin V/PI双染观察各组细胞凋亡情况;Western blot检测各组细胞中Cyclin B1、组蛋白H3的表达及其磷酸化水平的改变,以及凋亡相关蛋白Bcl-2、Bax、PARP表达的变化。结果:免疫荧光检测发现1μmol/L AZD1152作用MDA-MB-231细胞48 h时可见到多核细胞形成;MTT试验结果显示对照组、照射组、抑制组和联合组的细胞增殖抑制率分别为(0.61±0.32)%、(17.62±1.41)%、(29.67±0.41)%、(53.17±1.26)%;CCK-8检测显示细胞存活率分别为(94.88±0.22)%、(59.21±0.14)%、(42.05±0.17)%、(32.12±0.36)%;细胞周期检测发现G2/M期占比分别为(18.99±0.15)%、(38.05±0.23)%、(49.80±0.32)%、(75.52±0.45)%,与对照组比较差异均具有统计学意义(P<0.05)。照射组、抑制组和联合组的细胞凋亡率分别为(17.48±0.24)%、(29.23±0.02)%、(63.11±0.27)%,均较对照组(0.31±0.03)%升高(P<0.05)。流式细胞术发现抑制组细胞出现多核及多倍体细胞,易形成非整倍体。Western blot检测结果显示联合组较其他3组,抗凋亡蛋白Bcl-2、组蛋白H3的磷酸化和Cyclin B1蛋白表达减少(P<0.05),而促凋亡蛋白Bax和PARP蛋白剪切明显增加(P<0.05)。结论:^(125)I粒子对AZD1152有增敏作用,^(125)I联合AZD1152可显著抑制MDA-MB-231细胞组蛋白H3磷酸化及Cyclin B1水平,从而抑制细胞增殖,诱导细胞凋亡。OBJECTIVE:To study effects of AZD1152 combined with^(125)I particles on proliferations and apoptoses of triple negative breast cancer cells MDA-MB-231.METHODS:Cultured cells were divided into several groups;control,^(125)I particle irradiation,1μmol/L AZD1152 treatment with 48 h,treated with^(125)I particle irradiation and 1μmol/L AZD1152 treated with 48 h.Cell proliferation inhibitory rates were detected using the MTT and cell viability using the CCK-8 assays.Cell ploidies,cell cycles and apoptoses were detected using flow cytometry with cells separately stained by propidium iodide(PI)and by Annexin V/PI double-staining.Expressions of apoptosis-related proteins(Bcl-XL,Bcl-2 and PARP),CyclinB1 and Phosphorylation levels of Histone H3 were analyzed using Western blotting.RESULTS:After treatments for 48 h,cell proliferation inhibitory rates were(0.61±0.32)%,(17.62±1.41)%,(29.67±0.41)%,(53.17±1.26)%,respectively.Cell viability rates were(94.88±0.22)%,(59.21±0.14)%,(42.05±0.17)%(32.12±0.36)%,respectively.The proportions of G2/M phase were(18.99±0.15)%,(38.05±0.23)%,(49.80±0.32)%,(75.52±0.45)%,respectively.The apoptosis rates of MDA-MB-231 cells were(17.48±0.24)%,(29.23±0.02)%,(63.11±0.27)%,with significantly differences in the different groups(P<0.05).Compared with(0.31±0.03)%in the control group,the observed increases were significant(P<0.05).Immunofluorescence and flow cytometry analyses showed that multinucleated and polyploid cells appeared in the 1μmol/L AZD1152 group in 48 hours,which were easy to form aneuploid cells.Compared with the irradiated,inhibitor and control groups,the combined irradiation with 1μmol/L AZD1152 for 48 h effectively down-regulated the expressions of Bcl-2,p-Histone H3,CyclinB1 and promoted Bax and dissection of PARP(P<0.05).CONCLUSION:The combination of^(125)I particles and AZD1152 treatments efficiently inhibited proliferation and promoted apoptosis in breast cancer cell line MDA-MB-231.

关 键 词：乳腺癌 ^(125)I粒子 AURORA激酶 细胞增殖 凋亡 

分 类 号：R737.9[医药卫生—肿瘤]