ZNF300促进巨核细胞分化

ZNF300 Promotes Megakaryocytic Differentiation

目的:研究ZNF300对巨核细胞分化和增殖的影响。方法:通过公共数据分析及定量RT-PCR方法检测ZNF300表达水平以研究ZNF300与白血病及巨核细胞分化的相关性;使用慢病毒和逆转录病毒感染介导过表达ZNF300,使用流式细胞术、MTT比色法、集落形成试验分析K562细胞和原代小鼠骨髓细胞向巨核细胞分化、增殖的情况;应用细胞浆和细胞质蛋白分离结合蛋白印迹实验方法检测ZNF300亚细胞定位;并应用双荧光素酶试验和用ChIP-qPCR技术探讨ZNF300调控的靶基因。结果:ZNF300表达水平伴随巨核细胞的分化而升高;TPA诱导后巨核细胞分化的表面标记CD41、CD61在过表达ZNF300的K562细胞明显比对照细胞增多。同时,ZNF300显著降低K562细胞增殖率、阻滞G1ME细胞周期并降低小鼠骨髓细胞集落形成单位。ZNF300蛋白主要位于细胞核内,能够结合在C-MYC基因的启动子区并增强启动子活性。结论:过表达ZNF300可促进巨核细胞分化。

Objective： To study the function of ZNF300 in the megakaryocytes differentiation and proliferation.Methods： Public data analysis of ZNF300 expression and megakaryocyte culture were used to reveal the correlation of ZNF300 expression with leukemia and megakaryocyte differetniation; ZNF300 overexpression was mediated by lentiviral or retroviral infection,and the differentiation and proliferation of K562 cells and primary mouse bone marrow cells to magekaryocytes were measured by flow cytometry,MTT assay and colony-forming test; the ZNF300 subcellular localization was tested by separating cytosolic and nuclear extracts combined with Western blotting. The dual-luciferase assay and ChIP-qPCR were used to study ZNF300 target gene. Results： ZNF300 expression upregulation correlated with megakaryoyte differentiation; over-expression of ZNF300 promoted CD41 and CD61 expression,inhibited cell cycle progress,and could reduce colony-forming unit. The ZNF300 locolized in nuclear and regulated C-MYC expression.Conclusion： ZNF300 promotes megakaryocyte differentiation.