摘要
目的:研究华蟾素与拓扑异构酶Ⅱ抑制剂依托泊苷(VP16)诱导人单核细胞白血病细胞凋亡的协同作用。方法:U937细胞分别经0.225μg/ml华蟾素、10μg/mlVP16及两药联合作用1～5d。甲基噻唑基四唑(MTT)法检测细胞生存率,荧光染色观察细胞凋亡的形态改变,凝胶电泳观察DNA片段化改变。流式细胞仪定量分析细胞凋亡。结果:作用1d华蟾素组有抑制作用(P=0.014),而VP16组与合用组几乎没有抑制作用(P>0.05);作用2d,单药组均有明显抑制作用(P≤0.01),合用组也有抑制作用(P=0.048);作用4d后,各组均有明显抑制作用(P=0.000)。作用2d,各组U937细胞出现凋亡的形态改变,DNA电泳可见“梯子”条带。14例原代细胞分别经各组作用2d,合用组凋亡率较单药组高(P=0.025)。结论:华蟾素及VP16能抑制U937细胞生长及诱导凋亡。两药合用能增强诱导部分原代细胞凋亡的作用。
Objective To study the apoptosis of human monocytic leukemia cells induced by topoisomerase H inhibitor etoposide （VP16） with the cooperation of cinobufacini-induced. Methods U937 cells were treated respectively by 0.225 μg/ml cinobufacini,10 μg/ml VP16,or simultaneous use of the two drugs for 1-5 d. The cell viability was measured with methyl thiazolyl tetrazolium（MTT） assay. The morphological changes were observed through immunofluoresence staining. The DNA fragment was visualized by agarose gel electrophoresis. The apoptosis rate was analyzed by flow cytometry. Results In comparison with the corresponding treatment groups,U937 cells were inhibited by cinobufacini after 1 d （P= 0.014） ,but the cells were not inhibited by VP16 or simultaneous use of the two drugs （P〉0.05）. After 2 d,U937 cells were significantly inhibited by either of the drugs（P≤0.01 ） and simultaneous use of the two drugs（P=0.048）. After 4 d, U937 cells were significantly inhibited in all the groups（P=0.000）. After 2 d ,the morphological changes indicated apoptosis. The DNA eletrophoresis revealed the ＂ladder＂ pattern. The primary cells from 14 patients with acute monocytic leukemia were treated by all the above-mentioned treatments for 2 d. The apoptosis rate of the primary cells induced by simultaneous use of the two drugs was higher than that induced by either of the drugs（P=0.025）. Conclusions Cinobufaeini and VP16 could inhibit the proliferation of U937 cells and induce apeptosis. The apeptosis of the primary cells from some patients is enlarged by the simultaneous use of cinobufacini and VP16.
出处
《内科理论与实践》
2007年第2期114-117,共4页
Journal of Internal Medicine Concepts & Practice
基金
国家教育部骨干教师课题项目(编号:GGJS200013)
