高三尖杉酯碱治疗急性白血病机制的研究

目的探讨高三尖杉酯碱（HHar）对急性髓细胞白血病（AML）的疗效明显优于急性淋巴细胞白血病（ALL）的可能机制.方法以成人急性白血病（AL）原代细胞为研究对象,充分利用其遗传性和体内细胞相似的特点,模拟人体用药后的最大血药浓度,采用高、中、低3种浓度梯度对HHar进行研究.首先采用光镜、扫描电镜、透射电镜、DNA凝胶电泳、二苯胺法、流式细胞术等一系列国内外检测细胞凋亡的经典方法,充分证实HHar确可诱导AL原代细胞凋亡.尔后,进一步选取其中较为经济的光镜下计数细胞凋亡率、二苯胺法定量测定凋亡细胞的DNA片段化率及锥虫蓝染色光镜下计数坏死率3种方法定量观察HHar对ALL及AML原代细胞的诱导凋亡和杀伤,得出相应的时间、剂量和效应关系,并进行对比分析.结果HHar可诱导AL原代细胞凋亡,AML原代细胞对HHar诱导凋亡的敏感性高于ALL原代细胞.结论AML原代细胞对HHar诱导凋亡的敏感性高于ALL原代细胞,这是其临床疗效优于ALL的可能机制.

The therapeutics mechanism of HHar in treatment of acute leukemia

Objective To appraise the possible mechanism of Hhar which is more effective on AML than for ALL.Methods Primary acute leukemia cells of 34 patients (AML 19, ALL 15) were treated with HHar in 3 concentration gradients. Light microscopy, SEM and TEM, DNA agarose gel electrophoresis, diphemylamine reaction and cytometry were performed to prove that HHar could induce primary acute leukemia cells apoptosis. Then, the apoptotic and necrosis rates were calculated under the light microscope respectively and the internucleosomal DNA fragmentation ratios by diphemylamine reaction of the primary acute leukemia cells treated with HHar were measured. Finally, the time-dose-effect relationships were analyzed.Results HHar could induce the apoptosis of primary ALL cells. And the AML primary cells were more sensitive to HHar inducing apoptosis than the ALL ones.Conclusion The AML primary cells are more sensitive to the HHar inducing apoptosis than the ALL ones. These may be the possible mechanism why HHar is more effective on AML than for ALL.