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为了研究2-甲氧基雌二醇(methoxyestradiol,2-ME)诱导骨髓增生异常综合征-难治性贫血伴原始细胞增多型(MDS-RAEB)细胞株MUTZ-1细胞凋亡的机制,将不同浓度的2-甲氧基雌二醇分别与MUTZ-1细胞在体外培养,同时设二甲亚砜和空白对照组。采用四甲基偶氮唑蓝(MTT)比色法测定2-ME对MUTZ-1细胞的生长抑制率,瑞氏-姬姆萨染色后观察2-ME引起细胞的形态学改变,流式细胞术分析细胞周期和凋亡率的变化,贝克曼全自动生化分析仪(synchron clinical system LX20)检测培养上清液中乳酸脱氢酶(lactate dehydrogenase,LD)的变化,DNA凝胶电泳验证2-ME诱导的细胞凋亡。结果表明:2-ME对MUTZ-1细胞的增殖具有明显的抑制作用,该细胞凋亡率明显升高,并呈现时间和剂量依赖性,经统计学处理与对照组相比较有显著性差异(P<0.05)。4μmol/L2-ME作用MUTZ-1细胞12小时后,细胞呈现典型的凋亡细胞形态特征;2-ME作用24小时后MUTZ-1细胞出现G2/M期阻滞;培养上清液中LD含量与对照组相比明显升高,差异具有显著性(P<0.05);4μmol/L2-ME作用MUTZ-1细胞48小时后,DNA凝胶电泳可见明显的DNA梯形条带。结论:2-ME对骨髓增生异常综合征细胞株MUTZ-1有较强的抗肿瘤效应,可能与细胞G2/M期阻滞引起的细胞凋亡有关;2-ME是一种有发展潜力的治疗骨髓异常综合征的药物。
To investigate the mechanism of 2-methoxyestradiol (2-ME) -induced apoptosis in MUTZ-1 cells of myelodysplastic syndrome-refractory anemia (MDS-RAEB) cell line, Different concentrations of 2-methoxyestradiol and MUTZ-1 cells were cultured in vitro, while dimethyl sulfoxide and blank control group. The inhibitory rate of 2-ME on the growth of MUTZ-1 cells was determined by MTT assay. The morphological changes of 2-ME-induced cells were observed after Wright-Giemsa staining. Flow cytometry The changes of cell cycle and apoptosis rate were analyzed. The changes of lactate dehydrogenase (LD) in the culture supernatant were detected by Beckman automatic biochemistry analyzer (synchron clinical system LX20). The DNA 2- ME-induced apoptosis. The results showed that 2-ME significantly inhibited the proliferation of MUTZ-1 cells, and the apoptosis rate of MUTZ-1 cells was significantly increased. The apoptosis rate of MUTZ-1 cells was significantly increased in a dose-and time-dependent manner. Compared with the control group, P <0.05). After treated with 4μmol / L 2-ME for 12 hours, MUTZ-1 cells showed typical morphological features of apoptotic cells; G2 / M arrest occurred in MUTZ-1 cells after 2-ME treatment for 24 hours; LD in cultured supernatant (P <0.05). MUTZ-1 cells treated with 4μmol / L 2-ME showed obvious DNA ladder after DNA gel electrophoresis for 48 hours. Conclusion: 2-ME has a strong anti-tumor effect on myelodysplastic syndrome cell line MUTZ-1, which may be related to cell apoptosis induced by G2 / M arrest. 2-ME is a promising anti-tumor agent Treatment of myelodysplastic syndrome drugs.