转染外源性Fas配体基因的正常骨髓CD34^+细胞诱导U937细胞凋亡的实验研究

The Study of Apoptosis of Leukemia Cells Induced by CD34^+ Cells Transferred with Exogenous Fas Ligand

目的:观察转染外源性FasL的CD34+细胞对白血病细胞系U937的凋亡诱导作用,探讨通过向CD34+细胞转染FasL以增强移植后GVL效应的可能性。方法:免疫磁珠法分选骨髓CD34+细胞并进行体外扩增培养;FasL基因逆转录病毒途径转染CD34+细胞,并与白血病细胞U937混合培养,加入或不加入柔红霉素、阿糖胞苷,借助TUNEL、流式细胞仪检测细胞的凋亡率。结果:逆转录病毒上清转染后48～72h,流式细胞仪检测24.2%±2.4%CD34+细胞表达FasL(P<0.01)(未转染的CD34+细胞FasL阳性率为7.6%±1.1%)。未转染或转染外源FasL的CD34+细胞与白血病细胞U937以1:1的比例混合培养18h后,细胞凋亡率分别为5.0%±1.3%、10.8%±0.6%(P<0.01);FasL+CD34++DNR或FasL+CD34++Arac作用后,细胞凋亡率分别为13.4%±1.0%(P<0.05)、17.95%±1.3%(P<0.01)。结论:表达外源性FasL的骨髓CD34+细胞可诱导白血病细胞U937凋亡。提示植入转染外源性FasL基因的骨髓CD34+细胞可增强BMT后的GVL效应。

Objective: To assess the value of CD34+ cells transferred exogenous Fas ligand in inducing the apotosis of U937 cells. Methods: The CD34+ cells transfected with FasL or not- was mixed with leukaemic cells U937 with presence or absence of DNR or Ara-C. After18h- apoptosis of cells was detected by TUNEL and FCM. Results: Induced for 18h by CD34+ cells transfected with FasL or not- the ratio of apoptosis of U937 was 10.8%±0.6% (P<0.01)? 5.0%±1.3% respectively. Induced by FasL+CD34++DNR? FasL+ CD34++Arac- that was 13.4%±1.0% (P<0.05)? 17.9%±1.3% (P<0.01). Conclusion: CD34+ cells that was transfected with exogenous Fas ligand could induce apoptosis of human leukaemic cells- suggesting it was possible to enhance the graft-versus-leukemia effect by transplanting CD34+ cells transferred exogenous Fas ligand.