免疫毒素CCL25-PE38抗急性T淋巴细胞白血病实验研究

Study of CCL25-PE38 immunotoxin in treatment of T-cell acute lymphoblastic leukemia (T-ALL)

摘要:

目的  探讨免疫毒素CCL25-PE38对急性T淋巴细胞白血病（T-ALL）细胞系MOLT4（天然高水平表达CCR9）的杀伤作用，以期为T-ALL的靶向治疗提供基础。方法  通过流式细胞术、共聚焦显微术检测CCL25-PE38与MOLT4 细胞的结合及内化，Transwell趋化小室检测其趋化能力，细胞增殖试剂WST-1检测其细胞杀伤能力，Annexin V-FITC/PI染色检测其凋亡诱导效应，建立SCID小鼠白血病细胞异种移植肿瘤（CCR9+肿瘤）模型检测CCL25-PE38的抑瘤效果。结果  WST-1检测结果显示 CCL25-PE38 能特异性杀伤MOLT4细胞；Annexin V-FITC/PI染色显示CCL25-PE38主要通过凋亡诱导效应引起MOLT4细胞死亡；在SCID小鼠白血病细胞异种移植瘤模型中，注入CCL25-PE38能缓解CCR9+肿瘤的生长。结论  CCL25-PE38通过凋亡诱导作用引起MOLT4细胞死亡，缓解异种移植CCR9+肿瘤的生长。

Abstract:

Objective To investigate the killing effect of CCL25-PE38 immunotoxin on MOLT4 cell line in T-cell acute lymphoblastic leukemia (T-ALL) (CCR9 natural high-level expression), and to provide the foundation of targeting treatment of T-cell acute lymphoblastic leukemia (T-ALL). Methods Flow cytometryn and confocal microscopy were used to detect the binding and internalization of CCL25-PE38 and MOLT4 cells. Transwell chemotaxis chamber method was used to detect the chemotaxis. Cell Proliferation Reagent WST and Annexin V-FITC/PI stain were used to detect killing ability of cells and the apoptosis induced effect respectively. And antitumous effect of CCI25-PE38 was detected by building the SCID leukemia cells in mice xenograft tumor (CCR9+ tumors) model. Results WST-1 test showed that CCL25-PE38 produce specific lethal effect on MOLT4 cell. Annexin V-FITC/PI stain showed that the cause of death of MOLT4 cells was induced by apoptosis of CCL25-PE38. Injection of CCL25-PE38 in SCID leukemia cells in mice xenograft tumor (CCR9+ tumors) model can retard growth speed of CCR9+ cancer. Conclusions The cause of death of MOLT4 is induced by apoptosis-inducing effect of CCL25-PE38, which can retard the growth of CCR9+ xenotransplanted tumors.