Changes of Cell Cycle and Differentiation of Human Leukemia Cell Line HL60 Induced by nm23-H1 Overexpression
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摘要: 目的 探讨nm23-H1基因过表达对人早幼粒白血病细胞株HL60细胞周期及分化的影响。方法 构建nm23-H1 cDNA的真核表达载体pEGFP-N1-nm23H1，脂质体介导瞬时转染HL60细胞；在荧光显微镜下观察nm23-H1-EGFP融合蛋白的表达情况；流式细胞术分析细胞周期变化；NBT还原比色法检测细胞的诱导分化能力。结果 转染48hnm23-H1-EGFP融合蛋白在HL60细胞内表达量最高，此时细胞周期处于S期的细胞明显增多，对分化诱导剂DMS0的敏感性下降。结论 nm23-H1基因的过表达能促进HL60细胞的增殖和抑制其分化。
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关键词:
- nm232H1 基因 /
- 白血病 /
- 细胞周期 /
- 细胞分化
Abstract: Objective To investigate effect s of nm23-H1 overexpression on cell cycle and differentiation of HL60 cells. Methods Recombinant expression vector pEGFP-N1-nm23 H1 was constructed and transfected into HL60 cells with the aid of liposome in a transient way. Expression of fusion protein nm23-H1-EGFP was detected under the fluorescence microscope. Flow cytomet ry was adopted to analyze cell cycle. Differentiation rate of the DMSO-induced HL60 cells was tested by NBT reduction assay. Results Expression level of recombinant plasmid pEGFP-N1-nm23 H1 in HL60 cells reached a peak at 48h after transfection. Meanwhile, proportion of S-phase cells increased dramatically and HL60 cells became less sensitive to differentiation inducer DMSO. Conclusion Overexpression of nm23-H1 gene could stimulate proliferation but inhibit differentiation of HL60 cells. nm23-H1 gene might be a potential target of leukemia treatment .-
Key words:
- nm23-H1 gene /
- Leukemia /
- Cell cycle /
- Cell differentiation
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