t(17;19)-急性淋巴细胞白血病细胞对肿瘤坏死因子相关凋亡诱导配体的敏感性及其意义  

Sensitivity of t(17;19) acute lymphoblastic leukemia cells to tumor necrosis factor-related apoptosis-inducing ligand and its significance

作　　者：张晓春[1] 郑晓敏[1] 陈诚[1] 李义德[2] 

机构地区：[1]宁夏医科大学总医院儿科,宁夏银川750004 [2]宁夏医科大学总医院医学实验中心,宁夏银川750004

出　　处：《吉林大学学报（医学版）》2014年第1期164-169,共6页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题(81060049)

摘　　要：目的:观察t(17;19)-急性淋巴细胞白血病(ALL)细胞对肿瘤坏死因子(TNF)相关凋亡诱导配体(TRAIL)所介导的细胞毒的敏感性及可能机制,并探讨其临床意义。方法:以t(17;19)-ALL细胞株4株、1例t(17;19)-ALL患者骨髓标本为实验组,其他ALL细胞株28株为对照组。流式细胞术测定细胞表面TRAIL受体表达;重组人可溶性TRAIL(rhsTRAIL)作用后,3 H-thymidine法测定其对细胞增殖的影响;FITC标记的Annexin-V染色及流式细胞术检测细胞早期凋亡;免疫印迹法观察细胞凋亡通路变化(caspase-8、Bia、caspase-3和PARP的表达)。结果:t(17;19)-ALL细胞表面死亡受体4(DR4)表达水平明显高于其他各组细胞株(P<0.05),死亡受体5(DR5)表达水平高于MLL+-ALL细胞株(P<0.05),诱骗受体1(DcR1)和诱骗受体2(DcR2)均呈阴性表达;rhsTRAIL浓度为100μg·L-1时,t(17;19)-ALL细胞抑制率接近100%,显著高于其他各组细胞株(P<0.05或P<0.01),该抑制作用在加入TRAIL中和抗体RIK-2及caspase广谱抑制剂z-VAD-fmk后被阻断;加入rhsTRAIL后,t(17;19)-ALL细胞发生早期凋亡,其凋亡率明显高于对照组细胞株(P<0.05);rhsTRAIL作用2h内,caspase-8、Bid、caspase-3和PARP出现活化条带。结论:t(17;19)-ALL细胞株对TRAIL高度敏感,并最终对移植物抗白血病(GVL)效应敏感,t(17;19)-ALL患者为获得长期存活,应及早进行同种造血干细胞移植(allo-SCT)。Objective To observe the sensitivity of t （17; 19） acute lymphoblastic leukemia （ALL） necrosis factor-related apoptosis-inducing ligand （TRAIL） -mediated apoptosis and to discuss its poss and clinical significance. Methods 4 t （17; 19） -ALL cell lines and a primary cell sample cells to tumor ible were used as experimental group and 28 other B-precursor ALL cell lines were used as control group. The cell surface expression of TRAIL receptors was detected by flow cytometry, and the inhibition of cell growth was measured by 3 H-thymidine uptake assay after treated with rhsTRAIL. Early apoptosis was detected by flow cytometry using Annexin-V staining and the changes of apoptosis signaling pathway after incubation with rhsTRAIL were checked by immunoblotting assay. Results The cell surface expression of death receptor 4 （DR4） in t （17; 19） - ALL cells was significantly higher than that in other cell lines （P〈0.05）. The relative fluorescence intensity （RFI） of DR5 was higher than 1.4 while it was lower than 1.3 in other cell lines. The cell growth of t （17; 19） -ALL was nearly 100% inhibited by rhsTRAIL at 100 μg · L-1 , which was much higher than other cell lines （P〈0.05 or P〈0.01）. The inhibition was blocked by RIK-2 （neutralizing antibody of TRAIL） and z-VAD-fmk （ inhibitor of caspase）, rhsTRAIL induced early apoptosis in t （17; 19） -ALL cells and the apoptotic rate was much higher than that in negative control （P〈0.05）. Cleaved bands of caspase-8, Bid, caspase-3 and PARP were observed within 2 h of incubation with rhsTRAIL. Conclusion t （17; 19） -ALL cells are highly sensitive to TRAIL and eventually to graft versus leukemia （GVL）. Allo-SCT should be performed in early stage to obtain long-term surviving in the patients with t （17; 19） -ALL.

关 键 词：t(17 19)-急性淋巴细胞白血病 肿瘤坏死因子相关凋亡诱导配体 移植物抗白血病 细胞凋亡 

分 类 号：R733.71[医药卫生—肿瘤]