CAJ | 학술논문

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；Westen blot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 MTT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)mol/L,ABT-737明显降低了VX-680在T47D中的IC50 (0.277±0.057) μmol/L,AB-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/L VX-680作用48 h出现多核现象；Western blot 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.
목적 관찰VX-680연합ABT-737대유선암세포T47D조망적영향.방법 상규배양T47D세포,용새서람(MTT)법검측VX-680연합ABT-737대유선암세포증식적영향；Westen blot법검측조망지표；면역형광염색급류식세포술관찰다핵현상；형광염색관찰세포조망형태학변화；류식세포술검측세포조망솔.결과 MTT법검측결과현시,VX-680단독용약대세포억제작용불명현,반수억제농도(IC50)위( 25.457±1.406)mol/L,ABT-737명현강저료VX-680재T47D중적IC50 (0.277±0.057) μmol/L,AB-737증가VX-680적세포억제작용정제량관계；ABT737 단약대세포억제작용불명현,IC50위(0.959±0.018) μmol/L,VX.-680여ABT737연합대세포적억제작용명현증가,IC50(0.268±0.072) μmol/L;면역형광염색결과현시1μmol/L VX-680작용48 h출현다핵현상；Western blot 검측취선감이린산핵당취합매(PARP)결과현시,여단독용약비교,VX-680여ABT737연합용약시PARP、Caspase-3전절명현증가,B세포림파류/백혈병-2기인(bcl-2)、bcl-xL표체감소、bax 표체증가,정제량화시간의뢰성.류식세포술현시VX-680연합ABT-737시T47D세포조망솔가체(46.40 ±0.41)％.결론 ABT-737가현저제고VX-680대유선암T47D세포적유도조망작용.
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 MTT 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 multinuclear 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.