Luo Wen-Da, Ren Chang-Ming, Zhu Min, Chen Bao-Guo, Li Bo-Li, Dai Mei-Zhen, Guo Qun-Yi
Department of Hematology, Taizhou Hospital of Zhejiang Province, Linhai 317000, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2005 Apr;13(2):278-81.
In order to study the influence of trichosanthin (TCS) on apoptosis and growth inhibition of human NB4 cells in vitro, the expression of annexin V and the change of DeltaPsim of NB4 cells induced by TCS was analyzed by FACS, and MTT assay was adopted to measure the growth inhibition ratio of NB4 cells treated with TCS. Apoptosis was assayed by agarose gel electrophoresis. The results showed the higher concentration of TCS and the longer the acting time, the stronger growth inhibition of NB4 cells. The expression of annexin V was positive, and the positive ratio was greatly enhanced with prolongation of acting time. DeltaPsim reduced gradually while the apoptosis cells increasing. DNA agarose gel electrophoresis showed a gradient, which confirmed that TCS could induce NB4 cells apoptosis. In conclusion, taken together, data show that TCS can inhibit NB4 growth in vitro, and induce apoptosis. Experiment provides an important evidence for application of TCS in clinical treatment of acute promyelocytic leukemia.
为研究天花粉蛋白(TCS)体外对人NB4细胞凋亡及生长抑制的影响,采用流式细胞术分析TCS诱导NB4细胞的膜联蛋白V表达及线粒体膜电位(ΔΨm)变化,采用MTT法检测TCS处理后NB4细胞的生长抑制率。通过琼脂糖凝胶电泳检测细胞凋亡情况。结果显示,TCS浓度越高、作用时间越长,对NB4细胞的生长抑制作用越强。膜联蛋白V表达呈阳性,且随着作用时间延长阳性率显著升高。ΔΨm逐渐降低,凋亡细胞增多。DNA琼脂糖凝胶电泳呈现梯状条带,证实TCS可诱导NB4细胞凋亡。综上所述,数据表明TCS能在体外抑制NB4细胞生长并诱导其凋亡。本实验为TCS在急性早幼粒细胞白血病临床治疗中的应用提供了重要依据。