CAJ | 학술논문

目的：研究生存素（ survivin）抑制剂YM155对三阴性乳腺细胞株MDA-MB-231的凋亡效应及其可能的作用机制。方法采用CCK-8法实验检测不同浓度YM155对MDA-MB-231细胞增殖的影响，计算其24及48 h半数抑制浓度（ the half maximal inhibitory concentration ，IC50）；流式细胞术检测细胞凋亡情况；RT-PCR法检测加药与对照组细胞的survivin 和bcl-2的mRNA的表达量；蛋白质印迹法检测sur-vivin、bcl-2、caspase-3、PARP蛋白表达变化情况。结果 YM155对MDA-MB-231具有明显的生长抑制效应且呈剂量和时间依赖性，24及48 h的IC50分别为（1.749±0.265）及（0.823±0.125） ng／ml。流式细胞仪检测显示YM155在0.5、1.0及1.5 ng／ml浓度对细胞的凋亡率分别是（10.93±0.94）％、（31.10±1.51）％及（46.83±2.92）％，与对照组（6.4±1.2）％相比差异具有统计学意义（P＜0.01）。 YM155可显著下调survivin的mRNA和蛋白表达，同时降低bcl-2和提高caspase-3、PARP的蛋白表达。结论 YM155能有效诱导乳腺癌细胞MDA-MB-231凋亡，其诱导凋亡机制是通过下调survivin蛋白，激活caspase凋亡通路，切割DNA而引发凋亡；bcl-2家族基因可能参与此过程。
목적：연구생존소（ survivin）억제제YM155대삼음성유선세포주MDA-MB-231적조망효응급기가능적작용궤제。방법채용CCK-8법실험검측불동농도YM155대MDA-MB-231세포증식적영향，계산기24급48 h반수억제농도（ the half maximal inhibitory concentration ，IC50）；류식세포술검측세포조망정황；RT-PCR법검측가약여대조조세포적survivin 화bcl-2적mRNA적표체량；단백질인적법검측sur-vivin、bcl-2、caspase-3、PARP단백표체변화정황。결과 YM155대MDA-MB-231구유명현적생장억제효응차정제량화시간의뢰성，24급48 h적IC50분별위（1.749±0.265）급（0.823±0.125） ng／ml。류식세포의검측현시YM155재0.5、1.0급1.5 ng／ml농도대세포적조망솔분별시（10.93±0.94）％、（31.10±1.51）％급（46.83±2.92）％，여대조조（6.4±1.2）％상비차이구유통계학의의（P＜0.01）。 YM155가현저하조survivin적mRNA화단백표체，동시강저bcl-2화제고caspase-3、PARP적단백표체。결론 YM155능유효유도유선암세포MDA-MB-231조망，기유도조망궤제시통과하조survivin단백，격활caspase조망통로，절할DNA이인발조망；bcl-2가족기인가능삼여차과정。
Objective To investigate the apoptosis induction effects and the possible mechanism of YM155 on triple negative breast cancer MDA-MB-231cells.Methods MDA-MB-231 cells were treated with dif-ferent concentrations of YM 155, and the survival rate of the cells was determined by CCK-8 assay and the half maximal inhibitory concentration ( IC50 ) value of YM155 was calculated .The apoptosis rate was examined by An-nexin V-FITC/PI double staining.mRNA expression of survivin and bcl-2 in MDA-MB-231cells was detected by RT-PCR.The protein expression of survivin , bcl-2, caspase-3, and PARP were detected by Western blot .Re-sults YM155 significantly inhibited the growth of MDA-MB-231 cells in a dose-and-time-dependenct way .IC50 was(1.749 ±0.265) ng/ml and(0.823 ±0.125) ng/ml respectively at 24 and 48 hours.The apoptosis rate of cells treated with 0.5 ng/ml, 1.0 ng/ml, and 1.5 ng/ml YM155 was (10.93 ±0.94)%,(31.10 ±1.51)%, and(46.83 ±2.92)%respectively, which had significant difference compared to that of the control group (6.4 ± 1.2)%(P<0.01).YM155 could significantly decrease mRNA and protein expression of surviving , besides, it reduced bcl-2 expression and increased caspase-3 and PARP protein expression .Conclusions YM155 can ef-fectively induce the apoptosis of MDA-MB-231 cells by downregulating survivin and activating caspase pathway . Bcl-2 might play a role in the apoptosis .