小鼠急性髓系白血病模型的构建

Construction of Mouse Acute Myeloid Leukemia Model

MLL基因的异常重排会引发急性淋巴系(ALL)和急性髓系白血病(AML)。该文详细阐述了利用逆转录病毒载体MLL-AF9构建小鼠AML模型的方法。该研究比较了免疫磁珠法与5-氟尿嘧啶(5-FU)方法富集骨髓细胞的效率,以及不同时间点收获的病毒对骨髓Lin-细胞感染效率的影响。通过流式检测发现, 5-氟尿嘧啶富集的骨髓Lin–细能够被48 h收获的病毒高效感染。受体鼠在移植了MLL-AF9感染的骨髓Lin–细胞60天后,外周血、骨髓、脾脏组织中均有大量的白血病细胞浸润, RT-qPCR也验证了白血病靶基因的表达上调,表明小鼠AML模型的成功构建。这项研究为从事白血病研究的科研人员提供了一种有效的小鼠急性髓系白血病模型,为研究白血病发病机理与研发白血病治疗药物提供有用的工具。

Abnormal rearrangements of MLL gene cause acute lymphoid(ALL)and acute myeloid leukemia(AML).Here,we describe a method of construction of a mouse AML model using a MLL-AF9 retroviral system.We compared the efficiencies of Lineage–(Lin–)bone marrow cells enrichent by using immunomagnetic beads and 5-fluorouracil(5-FU)treatment,and efficiencies of two different infection strategies.We found that 5-FU treatment of mice and 2-round of infection of Lin–bone marrow cells with viral supernatant 48 h post-transfection would lead to a highly efficient infection of MLL-AF9 retroviral vector.Moreover,sixty days after transplantation of MLL-AF9-infected Lin-bone marrow cells,leukemia cells were observed in the peripheral blood,and infiltration of leukemia cells were detected in the bone marrow and spleen of the recipients.In addition,RT-qPCR results verified a significant increase of transcription of MLL-AF9-targeted genes,further indicating an establishment of the AML mouse model.Our study provides a useful tool of studying the molecular mechanisms of leukemia pathogenesis and development of novel therapies.