【摘要】 目的：研究¹²⁵I 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）、¹²⁵I粒子照射组（照射组）、1 μmol/L AZD1152作用MDA-MB-231细胞48 h组（抑制组）、¹²⁵I粒子照射与1 μmol/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1 μmol/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：¹²⁵I 粒子对AZD1152有增敏作用，¹²⁵I 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。更多还原

【Abstract】 OBJECTIVE: To study effects of AZD1152 combined with¹²⁵I particles on proliferations and apoptoses of triple negative breast cancer cells MDA-MB-231. METHODS: Cultured cells were divided into several groups; control,¹²⁵I particle irradiation, 1 μmol/L AZD1152 treatment with 48 h, treated with¹²⁵I particle irradiation and 1 μmol/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）, Cyclin B1 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 1 μmol/L AZD1152 for 48 h effectively down-regulated the expressions of Bcl-2,p-Histone H3,Cyclin B1 and promoted Bax and dissection of PARP （P<0.05）. CONCLUSION: The combination of¹²⁵I particles and AZD1152 treatments efficiently inhibited proliferation and promoted apoptosis in breast cancer cell line MDA-MB-231.更多还原