硝普钠抑制K562 细胞增殖并诱导细胞凋亡的实验研究

目的: 观察外源性一氧化氮(NO) 供体硝普钠对K562 细胞增殖抑制及诱导细胞凋亡作用。方法: 将不同浓度的硝普钠与K562 细胞在体外培养, 观察其作用时间效应和剂量效应;用活细胞计数、MTT 法观察硝普钠对K562 细胞增殖的抑制作用;用DNA凝胶电泳、DNA 含量及细胞周期分析、Annexin-V/PI双标记和DNA 片段原位末端标记法等分析细胞凋亡。同时设高铁氰化钾(PFC) 对照组和空白对照组。结果: NO 能抑制K562 细胞生长, 并在一定的剂量范围内呈现作用时间和剂量的量效关系, 大部分细胞阻滞于G₀/G₁ 期;K562 细胞与硝普钠作用后出现典型的细胞形态改变, DNA 片断化, 亚G₁ 峰检出并显著增加, Annexin V/PI 和DNA 片段原位末端标记表达增加等均证实NO 能诱导K562 细胞凋亡。而对照组并无此类变化。结论: NO 通过阻滞G₀/G₁期细胞显著抑制K562 细胞的增殖, 并有很强的致凋亡作用。

Effects of sodium nitroprusside on proliferation inhibition and apoptosis in K562 cell line

AIM: To study the effects of nitric oxide donor sodium nitroprusside on proliferation inhibition and apoptosis in K562 human leukemia cell line.METHODS: The different concentration of sodium nitroprusside and K562 cell were cultivated at the different time in vitro.The proliferation inhibition was analyzed by MTT assay and alive cell count.Cell apoptosis was analyzed by cell morphology, DNA agarose gel electrophoresis, DNA content, and Annexin V/PI labeling method.The TDTmediated dUTP nick end labeling (TUNEL) assay was used to quantitate the cell apoptosis in situ.The PFC and the blank were used as controls.RESULTS: NO inhibited K562 cell proliferation within a certain range of treating time and dosage, and a majority of K562 cells were arrested in G₀/G₁ phase.The K562 cells apoptosis was confirmed by type cell morphology, DNA fragment, sub-G₁ phase, TUNEL and Annexin V/PI labeling method with a time and dosage relationship, and the control group had no change like this.CONCLUTION: NO can suppress proliferation of K562 cell line by arresting G₀/G₁ phase and trigger apoptosis of the line.