摘要
目的建立小鼠急性髓细胞白血病（AML）模型，探讨P13K／Akt／mTOR信号通路抑制剂PP242及酪氨酸受体抑制剂达沙替尼对AML发生发展的影响。方法用携带MSCV—MLL-AF9-IRES-GFP的逆转录病毒感染C57BL／6J小鼠骨髓Lin一细胞，再将细胞移植至致死剂量照射的小鼠体内，诱发白血病，建立小鼠AML模型。小鼠发病后分别用溶剂（对照组）、达沙替尼、PP242、达沙替尼＋PP242进行处理。观察小鼠的存活情况，并每隔4天利用流式细胞仪检测各组小鼠外周血、骨髓中白血病细胞生长情况，检测不同药物处理后白血病细胞的凋亡、细胞周期。利用流式分选的方法分选不同药物处理后AML小鼠中的白血病细胞，提取RNA并逆转录合成cDNA，用聚合酶链反应的方法检测相关基因的表达。利用基因测序寻找调控抑制白血病细胞生长的相关分子机制并进行功能验证。结果与对照组相比，PP242处理组或达沙替尼处理组小鼠的寿命均延长，但是同时用PP242和达沙替尼处理组小鼠的存活时间较单独用PP242处理组或达沙替尼处理组明显延长；且同时用PP242和达沙替尼处理的小鼠外周血及骨髓中白血病细胞比例更低，联合酪氨酸受体抑制剂达沙替尼可使更多的白血病细胞处于细胞周期的G0期，联合酪氨酸受体抑制剂达沙替尼可促使白血病细胞发生凋亡。结论P13K／Akt／mTOR信号通路抑制剂PP242联合酩氨酸受体抑制剂达沙替尼应用可促使更多的白血病细胞凋亡及使更多的白血病细胞处于细胞周期的GO期，明显延缓AML的发生发展，可能会为临床治疗提供一个更好的方案。
Objective To establish the acute myeloid leukemia （AML） mouse model, and to preliminarily investigate the efficiency of dasatinib, a tryosine kinase inhibitor, and PP242, an inhibitor of PI3K/Akt/mTOR signaling pathway in the development of AML. Methods The lineage- （ Lin- ） cells of C57BL/6J were transduced with retrovirus carrying MSCV-MLL-AF9-1RES-GFP fusion gene. The transduced cells were transplanted into lethally irradiated recipient mice to induce AML, and then the AML mouse model were established. The leukemia mice were treated with vehicle, dasatinib, PP242, dasatinib ＋ PP242, separately. The survival of the recipient mice was observed and the percentage of leukemia cells in peripheral blood （PB） and bone marrow （BM） was examined every four days. The apoptosis rates and cell cycle status of leukemia cells were also examined by FLOW. The leukemia cells in different group were sorted, the mRNA of these leukemia were extracted and reverse transcripted for related gene expression by qRT-PCR. The molecular mechanism of supression of leukemia cells growth was studied via RNA-Seq experiments. Results Compared with control group, either PP242 or dasatinib could prolong the survival rate of recipient mice, however, the combination treatment AML mice with PP242 and dasatinib prolonged the life span of AML mice more significantly. The combination of PP242 and dasatinib could decrease the percentage of leukemia cells in PB and BM more significantly, arresting more leukemia cells in GO phase, inducing more apoptosis of leukemia cells. Conclusion Combination of tryosine kinase inhibitor-dasatinib and PI3K/Akt/mTOR signaling pathway inhibitor- PP242 could delay the progression of AML by inducing more leukemia to apoptosis and arrest more leukemia cells in the cell cycle GO phase, it may be provied a new method for clinical therapy.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2017年第20期1584-1588,共5页
National Medical Journal of China
关键词
白血病
髓样
急性
小鼠
药理作用分子作用机制
Leukemia, myeloid, acute
Mice
Molecular mechanisms of pharmacological
