摘要
目的 :探讨阿克拉霉素对HeLa细胞染色体形成的影响机制及与细胞凋亡的关系。 方法 :MTT方法测定阿克拉霉素对HeLa细胞的IC50 值。不同浓度阿克拉霉素作用于HeLa细胞24h后 ,制备染色体 ,Giemsa染色 ,观察 ;应用吖啶橙和嗅乙啶双荧光染色及DNA凝胶电泳检测细胞凋亡 ;Western_blot分析半胱氨酸蛋白酶_3(caspase_3)含量变化。 结果 :阿克拉霉素对HeLa细胞的IC50 值为6.83μg/ml。染色体分析表明 ,阿克拉霉素作用于HeLa细胞一定时间后 ,染色体不规则凝集 ,正常M期染色体形成受阻。细胞凋亡检测可见10μg/ml药物组与0.1μg/ml组及对照组相比 ,差异显著。Western_blot分析表明 ,10μg/ml药物组caspase_3表达比0.1μg/ml组和对照组显著增加。 结论 :阿克拉霉素能够阻止HeLa细胞染色体形成与分离 ,导致细胞G2/M期阻滞 ,诱导细胞凋亡。
Purpose: To investigate the mechanism of chromosome formation of HeLa cell line induced by aclarubicin and the relationship of aclarubicin to apoptosis. Methods: MTT assay was used to detect the half_inhibition concentration (IC 50) of a clarubicin on HeLa cell line, then aclarubicin with different concentrations was chosen to treat HeLa cells for 24 hours, and the nuclei and chromosomes were stained with Giemsastain. Apoptosis of HeLa cell was detected with acridine orange and ethidium bromide(AO / EB) double fluorescence stain and agarose g el electrophoresis. Caspase_3 was assessed with western blot met hod. Results: After HeLa cells were treated with aclarubicin, the IC 50 was 6.83 μg / ml. The normal chromosome formation w as blocked and chromatin condensed disorderly. Apoptosis and DNA fragments were apparent in the 10 μg / ml group. The caspa se_3 content in the 10 μg / ml group also showed a marked increase compared with the control and the 0.1 μg / ml group . Conclusion: Aclarubicin can inhibit the normal chromatin conde nsation and induce the cells G 2 / M arrest, and causes apopto sis. Caspase_3 may play an important role in apoptosis of He La cells induced by aclarubicin.
出处
《癌变．畸变．突变》
CAS
CSCD
2003年第2期84-87,共4页
Carcinogenesis,Teratogenesis & Mutagenesis
