苯丙氨酸解氨酶诱导人白血病HL-60细胞凋亡的初步研究

Preliminary study on apoptosis of human leukemic HL-60 cells induced by phenylalanine ammonialyase

目的 :研究苯丙氨酸解氨酶(L phenylalanineammonia lyase ,PAL)对白血病HL 60细胞生长的抑制作用 ,探讨其作用机制。方法 :应用MTT比色法观察PAL对HL 60细胞的生长抑制作用 ,应用倒置显微镜和DNA琼脂糖凝胶电泳技术分析细胞凋亡。结果 :PAL与HL 60细胞共育时 ,能明显抑制细胞生长 ,半数抑制浓度 (IC50 )为 3 0 2U/mL ;细胞呈典型的凋亡形态学改变 ,细胞皱缩 ,染色质凝集 ,沿核膜内侧分布 ,可见新月形 ;细胞DNA裂解成 180～ 2 0 0bp及其倍数的片段 ,电泳得到特有的梯形条带 ,凋亡率与PAL剂量和作用时间呈依赖关系。结论 :PAL能抑制HL 60细胞的生长 ,诱导细胞凋亡是其抗肿瘤的作用机制之一。

OBJECTIVE: To study the mechanism of inhibiting effect of phenylalanine ammonialyase (PAL) on human leukemic HL 60 cells in vitro . METHODS:HL 60 cells were treated with PAL at different concentrations and the growth inhibition was analysed by MTT colorimetric assay. Cell apoptosis was evaluated by the converting microscope and agrose gel electrophoresis respectively. RESULTS: The treatment with PAL remarkably inhibited the growth of HL 60 cells. The IC 50 was 3 02 U/mL. Apoptosis of HL 60 cells was induced by PAL,showing the apoptosis changes in morphology, including cell shrinkage and chromatin agglutination. DNA ladder was observed with agrose gel electrophoresis. PAL induced the apoptosis of HL 60 cells in a dose time dependent manner.CONCLUSIONS:PAL can efficiently inhibit the growth of HL 60 cells. Induced apoptosis may be one of the mechanism for inhibitory effect of PAL on leukemia.