曲古菌素A和ATRA合用对PLZFRARα转基因阳性发病鼠骨髓细胞的诱导分化作用_医学论文.docVIP

曲古菌素A和ATRA合用对PLZF转基因阳性发病鼠骨髓细胞的诱导分化作用_医学论文 曲古菌素A和ATRA合用对PLZF转基因阳性发病鼠骨髓细胞的诱导分化作用_医学论文 作者：陈思宇，董颖，陈丽娟，张龙，王龙，陈强，王振义 　　【Abstract】 AIM: To investigate the effect of all METHODS: Molecular cloning technology was used to construct hCGPLZFRARα gene. HCGPLZFRARα transgenic mice were generated, in which PLZFRARα fusion gene was driven by hCG promoter to express in myeloid cells of mice. The genotype and phenotype were analyzed by PCR, Southern Blot, RTPCR, immunofluorescence, morphology of bone marrow cells, peripheral blood cells and spleen cells, pathology of bone, spleen and liver. Bone marrow leukemia cells were cultured in vitro. The change of cell morphology was observed by WrightGiemsa staining cell cycle distribution and expression of cell membrane surface differentiationrelated antigens (such as CD11b, CD117 and Gr1) were determined by flow cytometry assay. Expression of PLZF was analyzed by immunofluorescence. Functional differentiation was reflected by nitroblue tetrazolium(NBT) reduction ability. RESULTS:Some hCG PLZFRARα transgenic mice developed leukemia resembling human chronic myeloid leukemia. After treated with TSA and RA, bone marrow leukemia cells presented morphologically some features of differentiated cells the cell growth and proliferation were inhibited in a timedependent manner. The proportion of cells in S phage was decreased. The expression of PLZF relocated in treated cells. However, no significant difference in NBT assay was documented with the combination therapy. CONCLUSION: Combination of histone deacetylase inhibitor with ATRA can cause a partial differentiation of PLZFRARα positive cells. 　　【Keywords】 PLZF RA transgene histone deacetylase differentiation leukemia, promyelocytic, acute 　　【摘要】 目的： 研究组蛋白去乙酰化酶抑制剂曲古菌素A(trichostatin A,TSA)和ATRA对PLZF转基因发病鼠骨髓细胞的作用. 方法： 分子克隆技术构建hCG转基因构件. 显微注射建立仅在髓系细胞中表达PLZF的转基因小鼠模型. DNA，Southern blot，RT，免疫荧光，血象，骨髓象，脾细胞形态，病理等检测分析基因整合表达及发病情况. 取PLZF转基因发病鼠骨髓细胞体外培养，经TSA和ATRA作用一段时间后，Wright染色观察细胞形态，流式细胞仪检测细胞周期