摘要
目的探讨氨甲喋呤单体[L-（＋）MTX、D-（-）MTX]及其不同比例对映体混合体[（±）MTX,D∶L=1∶9-9∶1]对急性淋巴细胞白血病细胞系（BALL-1、CCRF-CEM）抑制作用的差异。方法用氨甲喋呤单体及其混合体作用于BALL-1和CCRF-CEM细胞,MTT比色法分析其细胞增殖抑制率;荧光显微镜观察细胞形态学变化;碘化丙啶（PI）单染流式细胞术检测细胞周期;DNA梯度电泳检测其细胞凋亡水平。结果与对照组相比,MTX对映体对BALL-1、CCRF-CEM细胞的抑制生长呈现浓度和时间依赖性,其抑制强度依次为（±）MTX（D∶L=1∶9-2∶8）〉L-（＋）MTX〉D-（-）MTX;细胞周期出现S期的阻滞,DNA梯度电泳检测出现凋亡条带;亲本BALL-1、L-（＋）MTX/BALL-1、D-（-）MTX/BALL-1间差异有统计学意义（P〈0.01）。亲本CCRF-CEM、L-（＋）MTX/CCRF-CEM、D-（-）MTX/CCRF-CEM的细胞周期间差异亦有统计学意义（P〈0.01）。结论 MTX对映体对BALL-1、CCRF-CEM细胞的增殖抑制作用具有化学结构的立体选择性。
Objective To investigate the different inhibitory effects of methotrexate（ MTX） enantiomers [including L-（ ＋） MTX,D-（-） MTX,（ ±） MTX（ D∶ L =1∶ 9 -9∶ 1） ]on the proliferation of lymphoid leukemia cell lines BALL-1 and CCRF-CEM. Methods BALL-1 and CCRF-CEM cells were affected by the monomer or mixed type of MTX enantiomer. The inhibitory rates of proliferation of the cells were determined by MTT method. The morphology of the cells was inspected by fluorescence microscope. The phases of cell cycle were assayed by propidium iodide staining and flow cytometry. The apoptosis levels were detected by electrophoresis loading DNA ladder. Results Compared with the control groups,MTX enantiomers inhibited the growth of BALL-1 and CCRF-CEM cells in timeand dose-dependent manner. The inhibitory efficacy of（ ±） MTX（ D∶ L = 1∶ 9 and 2∶ 8） was higher than those of both L-（ ＋） MTXand D-（-） MTX,and the inhibitory efficacy of L-（ ＋） MTX was higher than those of（ ±） MTX（ D∶ L = 3∶ 7,4∶ 6,5∶ 5,6∶ 4,7∶ 3, 8∶ 2 and 9∶ 1） and D-（-） MTX,The inhibitory efficacy of D-（-） MTX was the lowest. The cell cycles were arrested in S phase,and the apoptosis bands were found in DNA ladder electrophoresis. There were significant differences of the cell cycle among parental BALL-1,L-（ ＋） MTX / BALL-1and D-（-） MTX / BALL-1（ P〈0. 01）,or parental CCRF-CEM,L-（ ＋） MTX / CCRF-CEM and D-（-） MTX/CCRF-CEM（ P〈0. 01）. Conclusion The inhibitory effects of MTX enantiomers on proliferation of BALL-1 and CCRFCEM cells presented chiral-selective in chemical structure.
出处
《临床检验杂志》
CAS
CSCD
北大核心
2014年第5期378-381,共4页
Chinese Journal of Clinical Laboratory Science
基金
国家自然科学基金(30672011)
