Aurora激酶抑制剂与BH3模拟剂协同诱导乳腺癌细胞凋亡  

The combination of VX-680 and ABT-737 efficiently promotes breast cancer cell line apoptosis

作　　者：张月[1] 张斌[1] 冯炜红[1] 李媛媛[1] 曹旭晨[1] 

机构地区：[1]天津医科大学附属肿瘤医院乳腺一科乳腺癌防治教育部重点实验室天津市“肿瘤防治”重点实验室,300060

出　　处：《中华实验外科杂志》2012年第5期793-796,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81001186）;天津市自然科学基金资助项目（10JCYBJC14100）

摘　　要：目的观察VX-680联合ABT．737对乳腺癌细胞T47D凋亡的影响。方法常规培养T47D细胞，用噻唑蓝（MTT）法检测VX-680联合ABT-737对乳腺癌细胞增殖的影响；Westenblot法检测凋亡指标；免疫荧光染色及流式细胞术观察多核现象；荧光染色观察细胞凋亡形态学变化；流式细胞术检测细胞凋亡率。结果MTF法检测结果显示，VX-680单独用药对细胞抑制作用不明显，半数抑制浓度（IC50）为（25．457±1．406）μmol／L，ABT-737明显降低了VX-680在T47D中的Ic50（0．277±0．057）μmol／L，ABT-737增加VX-680的细胞抑制作用呈剂量关系；ABT737单药对细胞抑制作用不明显，IC50为（0．959±0．018）μmol／L，VX-680与ABT737联合对细胞的抑制作用明显增加，IC50（0．268±0．072）μmol／L；免疫荧光染色结果显示1μmol／LVX-680作用48h出现多核现象；Westernblot检测聚腺苷二磷酸核糖聚合酶（PARP）结果显示，与单独用药比较，VX-680与ABT737联合用药时PARP、Caspase-3剪切明显增加，B细胞淋巴瘤／白血病-2基因（bcl-2）、bcl-xL表达减少、bax表达增加，呈剂量和时间依赖性。流式细胞术显示VX-680联合ABT-737时T47D细胞凋亡率可达（46．40±0．41）％。结论ABT-737可显著提高VX-680对乳腺癌T47D细胞的诱导凋亡作用。Objective To study the effect of VX-680 combined with ABT737 on apoptosis of breast cancer cell line T47D. Methods T47D cells were cultured in vitro, and cells in logarithmic growth phase were used for this experiment. Cell proliferation was measured by methyl thiazol tetrazolium （MTT） assay. The expression of apoptosis related protein was detected by using Western blotting. The variety of nucleus was tested by using immunofluorescence. Morphological change of apoptotic cells were observed under the fluorescent microscopy. The apoptosis rate was examined by flow cytometry. Results The MTr assay showed that as a single agent, 50% inhibitory concentration （ IC50 ） of VX-680 was （25.457±1. 406） μmol/L. ABT-737 significantly decrease the IC50 of VX-680 to （0. 277±0. 057）μmol/L. ABT-737 didn＇t inhibit the proliferation of T47D cells with IC50 being （0. 959±0. 018） μmol/L, but enhanced the inhibitory effect of VX-680 in a does-dependent with IC50 being （0. 268±0. 072）μmol/L. VX-680 induced muhinuclear phenomenon tested by immunofluorescence. VX-680 combined with ABT737 induced apoptosis in a dose-and time-dependent manner tested by Western blotting. Compared with the single agent, combined use of VX-680 and ABT-737 accelerated the cleavage of poly （ADP-ribose） polymerase （PARP） and caspase3 , decreased the expression levels of bcl-2 and bcl-xL, and promoted the expression level of bax. Flow cytometry revealed that the combination of VX-680 and ABT-737 significantly induced apoptosis and increased apoptosis rate to （46. 40±0. 41 ） %. Conclusion The ABT-737 can significantly enhance VX-680-induced apoptosis of T47D cells.

关 键 词：乳腺癌 VX-680 ABT-737 脱噬作用 

分 类 号：R737.9[医药卫生—肿瘤]