CAJ | 학술논문

目的探讨氨甲喋呤[(±)MTX]及其对映体[(+)MTX、(-)MTX]对ECV304细胞的生长抑制作用并探讨其机制.方法采用培养的ECV304细胞,应用MTT比色法分析其活性;用光学显微镜观察细胞的形态学变化;碘化丙啶(PI)单染流式细胞术检测细胞周期.结果在0.1～150 μmol·L~(-1)范围内,(+)MTX、(-)MTX和(±)MTX作用于ECV304细胞24、48、72 h,均抑制细胞ECV304增殖,但抑制强度依次为(+)MTX>(±)MTX>(-)MTX,倒置显微镜观察不同浓度(±)MTX、(+)MTX和(-)MTX作用ECV304细胞不同时间后,出现细胞体积变小、核固缩等形态学改变.用10 μmol·L~(-1)的(+)MTX、(-)MTX和(±)MTX作用ECV304细胞48 h后,PI单染流式细胞术检测ECV304细胞周期的影响,表明MTX消旋体及单一体干扰ECV304细胞DNA合成.结论 (+)MTX和(-)MTX对ECV304细胞的抗增殖作用具有化学结构的立体选择性,(+)MTX的抗ECV304细胞增殖作用明显强于(-)MTX.
목적 탐토안갑첩령[(±)MTX]급기대영체[(+)MTX、(-)MTX]대ECV304세포적생장억제작용병탐토기궤제.방법 채용배양적ECV304세포,응용MTT비색법분석기활성;용광학현미경관찰세포적형태학변화;전화병정(PI)단염류식세포술검측세포주기.결과 재0.1～150 μmol·L~(-1)범위내,(+)MTX、(-)MTX화(±)MTX작용우ECV304세포24、48、72 h,균억제세포ECV304증식,단억제강도의차위(+)MTX>(±)MTX>(-)MTX,도치현미경관찰불동농도(±)MTX、(+)MTX화(-)MTX작용ECV304세포불동시간후,출현세포체적변소、핵고축등형태학개변.용10 μmol·L~(-1)적(+)MTX、(-)MTX화(±)MTX작용ECV304세포48 h후,PI단염류식세포술검측ECV304세포주기적영향,표명MTX소선체급단일체간우ECV304세포DNA합성.결론 (+)MTX화(-)MTX대ECV304세포적항증식작용구유화학결구적입체선택성,(+)MTX적항ECV304세포증식작용명현강우(-)MTX.
Aim To investigate the effect of MTX(included(±)MTX,(+)MTX and(-)MTX)on the proliferation of ECV304 cells and to explore its mechanisms.Methods ECV304 cells were cultured.The cell proliferation was determined by MTT.The morphological changes were inspected by inverted microscope.Cell cycle phases were assayed by propidium iodide staining flow cytometry.Results ECV304 cells were treated with(+)MTX,(-)MTX and(±)MTX at 1～150 μmol·L~(-1) for 24,48,72 h.The results showed that the proliferation of ECV304 cells was significantly inhibited under different conditions.The order of the inhibited efficacy was(+)MTX>(±)MTX>(-)MTX.The morphology of ECV304 cells were changed by(+)MTX,(-)MTX and(±)MTX treatment,which included the cell shrinkage,chromatin condensation.After administration of 10 μmol·L~(-1) of(+)MTX,(-)MTX and(±)MTX for 48 h,the cell cycle phases were assayed by propidium iodide staining flow cytometry.The result showed DNA replication was interfered by(+)MTX,(-)MTX and(±)MTX treatment.Conclusions The proliferation of ECV304 cells has the chiral selective effects by(+)MTX and(-)MTX treatment,and the inhibition on ECV304 cells proliferation of(+)MTX is significantly stronger than that of (-)MTX.