CAJ | 학술논문

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Western blot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.Western blot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-XL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.
목적 탐토Bcl-2억제제ABT-737연합순박대유선암T47D세포주조망적영향.방법 상규배양T47D세포,채용사갑기우담서람(MTT)법검측ABT-737연합순박대T47D세포증식적영향,채용Western blot법검측조망상관단백적표체,채용형광염색법관찰T47D세포조망적형태학변화,채용류식세포의검측T47D세포적조망솔.결과 MTT법검측결과현시,ABT-737가명현강저순박재T47D세포중적IC50[유순박단약적(26.00±1.41) μmol/L강위연합용약적(13.00±1.11)μmol/L].ABT-737증가순박대T47D세포적억제작용,병정제량의뢰성,이ABT-737단약대T47D세포적억제작용불명현.Western blot검측다취ADP핵당취합매(PARP)적전절결과현시,ABT-737명현강저료순박유도T47D세포조망적농도,가쾌료조망유도시간,ABT-737대순박유도T47D세포조망적증민작용정제량의뢰성,단약대T47D세포적작용불명현.여단약조상비,ABT-737여순박연합시T47D세포중PARP화caspase3적전절명현증가,이Bcl-2、Bcl-XL화Bax적표체무명현변화.형광염색화류식세포술검측결과균현시,ABT-737연합순박시,T47D세포적조망솔명현증가.결론 ABT-737가현저제고순박대유선암T47D세포적조망유도작용.
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 MTT assay showed that ABT-737 significantly decreased the IC50 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 Bcl-2,Bcl-XL,and Bax.Both flow cytometry and fluorescence microscopy showed that ABT-737 combined with cisplatin 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.