鬼臼噻吩甙、高温联用体外诱导白血病细胞HL-60的凋亡及净化

Apoptosis of Leukemic HL-60 Cells Induced by the Combination of Hyperthemia and Vm 26 and Purging of the Leukemic Cells in Vitro

:【目的】探讨高温联用鬼臼噻吩甙(Vm26)诱导白血病细胞的凋亡及净化。【方法】13例急性白血病骨髓、15例正常骨髓和10例次HL60细胞株用形态学、DNA凝胶电泳、流式细胞仪及半固体集落培养技术,观察42℃60min高温或联用鬼臼噻吩甙(Vm26)诱导HL60细胞的凋亡其及体外净化白血病细胞的效果。【结果】①HL60细胞经42℃60min、42℃60min+85μmin/LVm26处理后48h流式细胞检测的凋亡细胞(subG1%)达高峰,分别为764%±78%、934%±36%,P<005。DNA凝胶电泳24～48h处理后出现典型的梯状图谱,而对于正常骨髓细胞经上述同样处理后subG1分别为186%±51%、401%±32%(与HL60细胞比较P<001)。②高温42℃60min联用Vm26体外净化白血病细胞集落(LCFU)、HL60的抑制率分别为992%±1.8%、100%。而正常粒巨噬细胞集落(GMCFU)的抑制率为495%±45%。【结论】高温42℃60min联用Vm26诱导HL60细胞的凋亡比对正常造血细胞的凋亡强。

Objective To investigate the differences in apoptosis between normal hematopoietic cells and leukemic cell line induced by combination of hyperthermia(HT) and Vm 26 and purging of the leukemia cells in vitro. Method The bone marrow cells of 13 cases acute leukemia, 15 cases healthy people and 10 cases HL 60, it was observed the percentage of apoptosis cells respectively by using morphological method, flow cytometry analysis and DNA electrophoresis as well as the in vitro purging efficiency, using colony forming cell culture. Results ① When cells were incubated 42 ℃ 60 min alone or combined 42 ℃ 60 min with Vm 26 for 48 h, Sub G 1% of HL 60 was 76 4%±7.8%,93.4%±3.6%, respectively (P<0 01). Sub G 1% of GM CFU was 18.6%±5.1%, 40.1%±3.2%, respectively (vs HL 60, P<0 01). DNA from HL 60 cells showed ladder form. ② After treatment with 42 ℃ 60 min plus Vm 26 (8.5 μml/L), inhibition rate was 99.2%±1.8% for L CFU, 100% for HL 60, 49.5%±4.5% for GM CFU. The HL 60 cells could be purged while the normal survived. Conclusion The effect of Vm 26 plus 42 ℃ 60 min induced apoptosis on HL 60 were stronger than those on normal hematopoietic cells. The combination of HT and Vm 26 was a good purging protocol of ABMT.