ABT-737对顺铂诱导乳腺癌T47D细胞凋亡的增敏作用  

Bcl-2 inhibitor ABT-737 enhances the cisplatin-induced apoptosis in breast cancer T47D cells

作　　者：陈祖锦[1] 张斌[1] 潘思虎[1] 赵洪猛[1] 张月[1] 冯炜红[1] 李媛媛[1] 曹旭晨[1] 

机构地区：[1]天津市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：《中华肿瘤杂志》2011年第12期891-895,共5页Chinese Journal of Oncology

基　　金：国家自然科学基金（81001186）;天津市自然科学基金（IOJCYBJC14100）

摘　　要：目的探讨Bcl-2抑制剂ABT-737联合顺铂对乳腺癌T47D细胞株凋亡的影响。方法常规培养T47D细胞，采用四甲基偶氮唑蓝（MTT）法检测ABT-737联合顺铂对T47D细胞增殖的影响，采用Westernblot法检测凋亡相关蛋白的表达，采用荧光染色法观察T47D细胞凋亡的形态学变化，采用流式细胞仪检测T47D细胞的凋亡率。结果MTT法检测结果显示，ABT-737可明显降低顺铂在T47D细胞中的IC50[由顺铂单药的（26．00±1．41）μmol／L降为联合用药的（13．00±1．11）μmol／L]。ABT-737增加顺铂对T47D细胞的抑制作用，并呈剂量依赖性，而ABT-737单药对T47D细胞的抑制作用不明显。Westernblot检测多聚ADP核糖聚合酶（PARP）的剪切结果显示，ABT-737明显降低了顺铂诱导T47D细胞凋亡的浓度，加快了凋亡诱导时间，ABT-737对顺铂诱导T47D细胞凋亡的增敏作用呈剂量依赖性，单药对T47D细胞的作用不明显。与单药组相比，ABT-737与顺铂联合时T47D细胞中PARP和caspase3的剪切明显增加，而Bcl-2、Bcl—X。和Bax的表达无明显变化。荧光染色和流式细胞术检测结果均显示，ABT-737联合顺铂时，T47D细胞的凋亡率明显增加。结论ABT-737可显著提高顺铂对乳腺癌T47D细胞的凋亡诱导作用。Objective To study the effect of ABT-737 combined with cisplatin on apoptosis of breast cancer cell line T47D cells. Methods T47D cells cultured in vitro was used for this experiment. Cell proliferation was measured by MTT assay. The expression of apoptosis-related protein was determined by Western blot. Morphological changes of apoptotic cells were observed by fluorescence microscopy. The apoptosis rate was examined by flow cytometry. Results The MTr assay showed that ABT-737 significantly decreased the IC5o of cisplatin in T47D cells [ （26.00 ± 1.41 ） μmol/L of single cisplatin vs. （ 13.00 ± 1.11 ） μmol/L of combination （ABT-737 ＋ cisplatin ）]. As a single agent, ABT-737 did not inhibite the proliferation of T47D cells, but enhanced the inhibitory effect of cisplatin in a dose-dependent manner. The detection of the cleavage of PARP showed that ABT-737 lowered the doses of cisplatin to induce apoptosis and shortened the induction time of apoptosis in T47D cells. Compared with the single use of cisplatin, the combination of ABT-737 and cisplatin accelerated the cleavage of PARP and caspase3, but did not alter the expression levels of Bel-2, Bcl-XL, and Bax. Both flow cytometry and fluorescence microscopy showed that ABT-737 combined with eisplatin significantly increased the apoptosis induction in T47D cells （2.3% ± 0.1% in the control, 30.0% ± 0.8% in the cisplatin alone, and 49.0% ± 0.5% in the cisplatin ＋ ABT- 737 groups, P 〈 0.05 ）. Conclusion The Bcl-2 inhibitor ABT-737 can significantly enhance cisplatininduced apoptosis in human breast cancer T47D cells in vitro.

关 键 词：乳腺肿瘤 ABT-737 顺铂 细胞凋亡 

分 类 号：R737.9[医药卫生—肿瘤]