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目的体外研究二乙酰己二胺(CAHB)对人骨髓增生异常综合征(MDS)细胞株MUTZ-1细胞的作用以及可能的作用机制。方法用CAHB处理MUTZ-1细胞,光学显微镜下观察不同浓度的CAHB处理后MUTZ-1细胞形态的改变;利用四甲基偶氮唑盐比色法(MTT法)检测CAHB对MUTZ-1细胞的增殖抑制作用;采用流式细胞仪分析细胞DNA含量分布变化;利用RT-PCR技术检测凋亡相关基因Survivin、Bcl-2的表达变化。结果CAHB能抑制MUTZ-1细胞的生长,具有浓度和时间依赖性;随着CAHB药物浓度的增加MUTZ-1细胞出现凋亡细胞的典型形态学特征改变;CAHB作用后,MUTZ-1细胞的DNA含量在细胞周期的分布有明显变化,G0/G1期细胞比例无明显变化,S期细胞减少,而G2/M期细胞增多,细胞被阻滞在G2期,呈浓度依赖;在细胞凋亡过程中抗凋亡基因Bcl-2、Survivin基因下调。结论CAHB不但能抑制MUTZ-1细胞生长而且能诱导细胞凋亡,诱导细胞凋亡是其主要细胞毒作用之一。抗凋亡基因Bcl-2、Survivin基因下调可能是其诱导细胞凋亡的机制之一。
Objective To investigate the effect of diacetylhexamethylenediamine (CAHB) on human myelodysplastic syndrome (MDS) cell line MUTZ-1 in vitro and its possible mechanism. Methods MUTZ-1 cells were treated with CAHB, and the morphological changes of MUTZ-1 cells were observed under light microscope with different concentrations of CAHB. MTT assay was used to detect the effect of CAHB on MUTZ-1 cells Proliferation inhibition; using flow cytometry analysis of cellular DNA content distribution changes; using RT-PCR detection of apoptosis-related genes Survivin, Bcl-2 expression changes. Results CAHB could inhibit the growth of MUTZ-1 cells in a concentration-and time-dependent manner. The typical morphology of apoptotic cells in MUTZ-1 cells was changed with the increase of CAHB concentration. After CAHB treatment, The content of cells in the cell cycle changes significantly, the proportion of G0 / G1 phase cells did not change significantly, S phase cells decreased, while G2 / M phase cells increased, the cells were arrested in G2 phase, in a concentration-dependent manner; in the process of apoptosis The anti-apoptotic genes Bcl-2 and Survivin were down-regulated. Conclusion CAHB can not only inhibit the growth of MUTZ-1 cells, but also induce apoptosis. Inducing apoptosis is one of the major cytotoxic effects. The anti-apoptotic gene Bcl-2, Survivin gene down regulation may be one of the mechanisms of its induction of apoptosis.