安吖啶诱导白血病细胞程序死亡及体外细胞药代动力学的研究

STUDIES ON AMSACRINE-INDUCED PROGRAMMED CELL DEATH AND IN VITRO CELLULAR PHARMACOKINETICS IN LEUKEMIA CELLS

本实验研究了ＮＢ４及ＨＬ－６０细胞的程序化死亡及细胞药代动力学特征。结果表明，安吖啶对ＮＢ４及ＨＬ－６０细胞均可诱导程序化死亡，但ＮＢ４细胞较ＨＬ－６０细胞更敏感，形态学改变更明显；ＤＮＡ凝胶电泳梯带更明显。安吖啶作用后，两株细胞的胞膜完整性可保持２４－４８ｈ。与４ｍｇ／Ｌ的安吖啶孵温后，非同步化细胞内安吖啶浓度在６ｈ达到高峰（ＮＢ４和ＨＬ－６０细胞分别为２５７．３９±３．２１ｎｇ／１０７细胞和１７４．９６±８．７６ｎｇ／１０７细胞），随后逐渐降低。但安吖啶诱导两株细胞程序化死亡的细胞数都随时间延长而逐渐增加。在３，６，１２ｈ时ＮＢ４细胞内药物浓度高于ＨＬ－６０细胞（Ｐ＜０．０１），Ｇ２Ｍ期的ＮＢ４细胞内的药物浓度较ＨＬ－６０细胞高（Ｐ＜０．０５）。安吖啶诱导程序化死亡作用与细胞内药物蓄积有关。

We studied PCD in NB4 and HL-60 cells and in vitro cellular pharmacokinetics of m-AMSA in synchronized and asynchronized cells. Our results showed that m-AMSA could induce PCD both in NB4 and HL-60 cell lines, and NB4 cells were more sensitive than HL-60 cells to m-AMSA:Typical morphological changes occured in NB4 cells were more evident than that in HL-60 cells. DNA ladders detected electrophoretically in DNA isolated from NB4 cells,was more evident than that in HL-60 cells.After exposure to m-AMSA, cellular membrane integrity could maintain for 24h-48 h in these two cell lines. Intracellular maximal concentration in two cell lines reached its peak in 6h 257.39±3.21 ng/107 cells in NB4 cells and 174.96±8.76 ng/107 cells in HL-60 cells, respectively, and then decreased gradually. However,PCD induced by m-AMSA was extensive and the number of PCD positive cells increased with prolongation of exposure time.Furthermore,there was significant difference between the two cell lines after incubation for 3h,6h,and 12 h(P<0.01). Intracellular m-AMSA concentration in G2M phase of NB4 cells was higher than that of HL-60 cells (P<0.05). Conclusion: m-AMSA-induced PCD correlated to cellular m-AMSA accumulation and there was a time-delayed reponse between them.