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)

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。

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.