重组人可溶性TRAIL分子诱导白血病细胞株凋亡的研究

比较重组人可溶性TRAIL(rhsTRAIL)诱导Jurkat细胞株、K562细胞株以及HL-60细胞株凋亡之间的差异,探讨这些差异与细胞表面TRAIL受体(DR4、DR5、DcR1和DcR2)表达量的关系。不同浓度的rhsTRAIL分别处理Jurkat细胞、K562细胞和HL-60细胞12 h、24 h和48 h后,用流式细胞仪检测经碘化丙啶(PI)染色后的细胞凋亡情况;用RT-PCR方法检测细胞表面受体DR4、DR5、DcR1、DcR2的表达。培养12 h、24 h、48 h后,不同浓度rhsTRAIL诱导Jurkat细胞株的凋亡率均明显高于对照组,且具有剂量依赖性和时间依赖性;但K562细胞株和HL-60细胞株未见明显的凋亡发生。RT-PCR结果显示,培养12 h、24 h、48 h后,Jurkat细胞株表面DR4的表达随时间的延长和rhsTRAIL浓度的升高而升高,而DR5、DcR1和DcR2的表达未检出;K562和HL-60细胞株表面DR4的表达没有明显变化,而且DR5、DcR1和DcR2的表达也未检出。rhsTRAIL诱导Jurkat细胞株的凋亡具有剂量依赖性和时间依赖性,且与其细胞表面DR4的表达呈正相关;在一定的浓度条件下,rhsTRAIL未能诱导K 562和HL-60细胞株发生明显凋亡,且其细胞表面DR4的表达也未见明显变化。这些结果提示,应用TRAIL治疗不同种类白血病时,应注意它的使用剂量和适应范围。

The difference in inducing apoptosis of leukemic cell lines by recombinant human soluble TNF-related apoptosis inducing ligand(rhsTRAIL)

To compare the difference in inducing apoptosis of leukemic cell lines Jurkat cells,K562 cells and HL-60 cells by recombinant human soluble TNF-related apoptosis inducing ligand(rhsTRAIL) and to explore the relationship between these differences and the expression quantity of the TRAIL receptors,such as DR4,DR5,DcR1 and DcR2,the leukemic cell lines employed were treated with different concentrations of rhsTRAIL for 12,24,and 48 hours respectively.The apoptosis was detected by PI staining and flow cytometry,and the expressions of the TRAIL receptors DR4,DR5,DcR1 and DcR2 were detected by RT-PCR.It was demonstrated that the apoptosis rate of Jurkat cells induced with different concentrations was higher than that of the untreated cells in a dose-and time-dependent manner.However,no significant evidence of apoptosis was found in case of the K562 and HL-60 cells line.As demonstrated by RT-PCR,the expression of DR4 on the surface of the Jurkat cells line was increased along with the prolongation of cultivation times for cells and the increasing concentrations of rhsTRAIL,but no expression of DR5,DcR1 and DcR2 could be detected.On the surface of the K562 cells and HL-60 cells,the expression of DR4 showed no significant change,and no expression of DR5,DcR1 and DcR2 could be detected also.The apoptosis of Jurkat cells induced by rhsTRAIL had dose-and time-dependence and showed positive correlation with the surface expression of DR4.It is evident that under certain concentration of rhsTRAIL induces no apoptosis of the K562 cells and HL-60 cells and no significant change on the surface expression of DR4 can be found.These results suggest that in case of treatment of patients with leukemia with TRAIL,its dosage and range of application should be carefully appreciated.