摘要
目的：构建bcr-abl逆转录病毒介导的小鼠慢性粒细胞白血病（CML）样模型,为深入研究CML发病机制和治疗奠定基础.方法：bcr-abl逆转录病毒Mig210载体经Phoenix-Ampo细胞包装后,取上清液感染5-FU处理后的BALB/c雄性小鼠骨髓细胞,再经尾静脉移植入经致死剂量γ射线（900cGy）照射的同种雌性受体鼠中.用形态学、RT-PCR和Western Blot鉴定小鼠成模情况.结果：小鼠移植8-9 wk后,外周血白细胞达2×10^10-3×10^10/L（为对照鼠的5-8倍）,外周血涂片幼稚细胞达到0.10-0.20;骨髓粒系细胞显著增高,易见幼稚细胞,肝脾也可见白血病细胞浸润;移植4-5 wk后在受体鼠骨髓和脾脏检出bcr-abl融合基因,8-9 wk后在肝脏亦检测出bcr-abl融合基因,同时在骨髓和脾脏也检测到bcr-abl融合蛋白.建模成功的小鼠3-5 mo后相继死亡.结论：成功建立bcr-abl逆转录病毒介导的小鼠CML模型,可用于后续CML信号通路和治疗机制的研究.
ATM： To construct the bcr-abl retroviral vectormediated mouse model of chronic myeloid leukemia like （CML-like） in order to establish a foundation for further investigation into its pathogenesis and therapeutic results. METHODS： The Mig210 retroviral vector containing the bcr-abl fusion gene was packaged by Phoenix-Ampo cells, and the retroviral supernatant, which was then used to infect the bone marrow cells from the 5- FU-treated male Balb/c donor mouse, was collected. After being infected by the retrovirus, the donor bone marrow ceils were transplanted into the lethally-irradiated （900cGyγ-ray） homogenous female receptor mouse intravenously via vena caudalis. Morphological observation, RT-PCR and Western Blot were employed to evaluate the mouse CML-like model system. RESULTS： After 8 -9 weeks, the white blood cells in the peripheral blood were significantly increased to 20 × 10^9 -30 × 10^9/L（5 - 8 times that of the normal control mice） ; and the proportion of the blast cells in the peripheral blood smear reached 10% - 20%. Results from bone marrow slides showed that the cell population of the granulocytic lineage also increased significantly and the blast cells were readily found. What＇s more, the leukemic cells were found to be infihrating into the liver and spleen of the candidate model mouse. The bcr-abl fusion gene was detected by RT-PCR in the bone marrow and spleen within 4 - 5 weeks, and in the liver within 8 - 9 weeks from the receptor mouse, and at the same time, the bcr-abl fusion protein was detected in the bone marrow and spleen as shown by Western Blot. The mice in which the CML-like model were successfully constructed died within 3 - 5 months. CONCLUSION： The bcr-abl retroviral vector-mediated mouse CML- like model is successfully constructed, which can be used in the subsequent researches on CML in the fields of cell signal transductions and therapeutic results.
出处
《第四军医大学学报》
北大核心
2008年第15期1377-1381,共5页
Journal of the Fourth Military Medical University
基金
国家自然科学基金(30470744
30670901)
