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目的:研究毛兰素对HL-60细胞增殖的抑制作用,探讨其诱导细胞凋亡的机制。方法:用MTT比色法测定了毛兰素对HL-60细胞增殖的抑制作用:应用荧光显微镜、透射电镜、DNA电泳及流式细胞仪观察了药物对细胞凋亡的诱导作用,并用免疫组化的方法从基因水平阐述了凋亡的发生。结果:毛兰素20-81.9nmol/L在72h内显著抑制HL-60细胞增殖,作用24h后,对HL-60细胞的IC_(50)为38nmol/L,而阳性对照药长春新碱对HL-60细胞的IC_(50)为101nmol/L,前者明显优于后者;形态学观察可见凋亡的特征性改变;琼脂糖电泳出现典型的DNA“ladder”;流式细胞仪结果表明细胞被阻滞于G_2/M期;免疫组化可见bcl-2表达下降,bax表达升高。结论:毛兰素显著抑制HL-60细胞的生长,该抑制作用可能是通过诱导细胞凋亡和改变HL-60细胞bcl-2和bax基因的表达而实现的。
OBJECTIVE: To study the inhibitory effect of Erianin on the proliferation of HL-60 cells and to explore the mechanism of its induction of apoptosis. METHODS: The inhibitory effect of Eriane on the proliferation of HL-60 cells was measured by MTT colorimetric assay: the induction of apoptosis by drugs was observed by fluorescence microscopy, transmission electron microscopy, DNA electrophoresis, and flow cytometry, and the immunization group was used. The method of genesis elucidates the occurrence of apoptosis from the gene level. RESULTS: Erythlineol 20-81.9 nmol/L significantly inhibited the proliferation of HL-60 cells within 72 h. After 24 h, the IC 50 of HL-60 cells was 38 nmol/L, and the positive control drug vincristine was HL. The IC50 of -60 cells was 101 nmol/L, the former was significantly better than the latter; morphological observations showed a characteristic change in apoptosis; typical DNA “ladder” appeared in agarose electrophoresis; flow cytometry results showed that the cells were Blocked in G 2 /M phase; immunohistochemistry showed that bcl-2 expression decreased, bax expression increased. CONCLUSION: Erythline inhibits the growth of HL-60 cells significantly. The inhibition may be achieved by inducing apoptosis and changing the expression of bcl-2 and bax genes in HL-60 cells.