K562细胞DANCE反应及其生化机制的研究.docVIP

  K562 细胞 DANCE 反应及其生化机制的 研究 吴彬彬，刘誉** 5 10 15 20 25 30 35 40 （暨南大学医学院生化教研室，广州 510632） 摘要：本课题研究 K562 细胞的 DANCE 反应并探讨其可能的生化机制。采用不连续蔗糖密 度梯度离心法(DSGC)将 K562 细胞分成不同条带，生化分析仪测定不同条带细胞内离子含 量，水分测定仪测定各条带细胞内水含量，流式细胞仪测定细胞的分裂周期及凋亡情况，实 验数据采用 SPSS12.0 软件进行统计分析。结果显示，DSGC 离心 K562 细胞得到 S50 和 S60 两条带，S50 条带 K、Ca 和 Mg 离子含量显著高于 S60 条带，同时 S50 条带水含量高于 S60 条带；药物作用后 S60 条带消失，甚至出现新的条带。流式结果显示 S60 条带的细胞凋亡 率最低，S30 的最高，提示细胞内密度减小凋亡增加。结论：细胞内离子含量增加伴随水含 量增加，导致细胞内密度下降，可能是 DANCE 反应的生化机制；药物作用后细胞内密度减 小，DANCE 上移反映了细胞的病理变化加重。 关键词：DANCE；不连续蔗糖密度梯度离心；水含量；细胞密度 中图分类号：Q503 Study of DANCE in K562 cells and its biochemical mechanism WU Binbin, LIU Yu (Deptartment of Biochemistry, Medical College,Jinan University, GuangZhou 510632) Abstract: The aim of the present study was to determine the DANCE response of K562 cells and to explore its biochemical mechanism. Discontinuous sucrose gradient centrifugation (DSGC) was used to separate K562 cells. Biochemical analyzer was used to determine the ion concentrations in the cells of indifferent bands, moisture analyser used to measure the water contents in the cells of different bands, and flowcytometer used to determine the cell cycle and appoptosis of different bands. The data were statistically analyzed by SPSS12.0 software. Our results showed that K562 cells were seperated into two bands, named S50 and S60 respectively. The contents of k、Ca and Mg in cells of S50 were higher than that of S60. The water content of S50 was higher than that of S60. After incubation with drugs, the band S60 disappeared, accompanied by the appearance of new bands. Flowcytometry found that cells of S60 were the lowest at appoptosis while S30 was the highest, indicating that intracellular density was reduced with apopptosis undergoing in the cells. We conclude that increase of intracellular ions accompanied by increase of water content in the cells might be the mechanism for decrease of intracellular density, and DANCE caused by drugs might reflex the pathological changes in the cells. Ke