环磷酰胺诱导白血病细胞株凋亡及FasL、FADD基因蛋白表达  

Cyclophosphamide-induced apoptosis and expression of FasL and FADD in leukemia cell line Jurkat

作　　者：刘家浩[1] 唐洪丽[1] 阮为勇[1] 王伟[1] 唐月华[1] 

机构地区：[1]东南大学附属中大医院儿科,南京210009

出　　处：《江苏医药》2006年第8期710-712,共3页Jiangsu Medical Journal

基　　金：江苏省社会发展项目(BS2003662)

摘　　要：目的研究环磷酰胺(CP)诱导白血病细胞株(Jurkat)凋亡的机制,进一步探讨FasL和FADD在CP诱导白血病细胞凋亡中的作用。方法CP刺激人T淋巴细胞白血病细胞株Jurkat,分别培养12、24、48h,在相应的时间点收获细胞,碘化丙啶(PI)染色细胞DNA,流式细胞术(FCM)检测DNA片段的亚二倍体峰(subG1)阳性的凋亡细胞;并用WesternBlot技术测定FasL和FADD的表达。结果CP3mg/L刺激Jurkat白血病细胞12h几乎未见细胞凋亡(P>0.05),在24h也只有26.40%的细胞出现凋亡;但随着时间的延长,CP诱导细胞凋亡明显增加,到48h则有74.95%的细胞发生凋亡(P<0.01)。CP也上调FasL和FADD的表达,并随CP刺激细胞时间的延长其表达量也逐渐增加。结论CP诱导白血病细胞株Jurkat凋亡的起效时间较晚,诱导细胞DNA片段形成,上调FasL和FADD的表达,表明CP诱导白血病细胞凋亡依赖于Fas/FasL通路。Objective To investigate the mechanism of apoptosis and expression of FasL and FADD induced by cyclophosphamide（CP） in leukemic cells. Methods After treated with CP,the Jurkat cell was cultivated for 12, 24 and 48 h, respectively. The apoptotic cells with subG1 peak were quantified by flow cytometry（FCM） as soon as the cells were stained with propidium iodide（PI）. The expression of FasL and FADD was detected by Western blot technique. Results There was almost no apoptotic cell after CP 3 mg/L stimulating the leukemic cells for 12 h（P〈0.05）. Only 26.40% of apoptotic cells were measured at 24 h, but as the time was prolonged, CP-induced apoptosis increased dramatically,and attained as high as 74. 95% at 48 h. The expression of FasL and FADD was detected after the cells were treated with CP, and the level of expression increased with stimulating time. Conclusion It took relatively a long time that CP induced apoptosis of leukemia cells. CP induced DNA fragmentation and upregulated the expresssion of FasL and FADD. It suggests that CP-induced apoptosis of leukemic cell is dependent on Fas/FasL pathway.

关 键 词：环磷酰胺 细胞凋亡 FASL FADD JURKAT细胞株 

分 类 号：R979.1[医药卫生—药品] R733.7[医药卫生—药学]