ZNF300促进巨核细胞分化

发布时间：2018-06-29 01:48

本文选题：ZNF基因 + 巨核细胞分化　；参考：《中国实验血液学杂志》2017年05期

【摘要】：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。
[Abstract]:Objective: to study the effect of ZNF300 on the differentiation and proliferation of megakaryocytes. Methods: the expression of ZNF300 was detected by public data analysis and quantitative RT-PCR in order to study the relationship between ZNF300 and leukemia and megakaryocyte differentiation, the overexpression of ZNF300 was mediated by lentivirus and retrovirus infection, and flow cytometry was used to detect the expression of ZNF300. Colony formation assay was used to analyze the differentiation and proliferation of K562 cells and primary mouse bone marrow cells into megakaryocytes, and the localization of ZNF300 subcells was detected by cytoplasmic and cytoplasmic protein separation and Western blotting assay. Double luciferase assay and ChIP-qPCR were used to study the target genes regulated by ZNF300. Results with the differentiation of megakaryocytes, the expression level of TPA-induced megakaryocyte CD41 + CD61 was significantly increased in K562 cells with overexpression of ZNF300. At the same time, ZNF300 significantly reduced the proliferation rate of K562 cells, blocked the cell cycle of G1ME cells and reduced the colony forming unit of mouse bone marrow cells. ZNF300 protein was mainly located in the nucleus, which could bind to the promoter region of C-MYC gene and enhance the promoter activity. Conclusion: overexpression of ZNF300 can promote megakaryocyte differentiation.
【作者单位】：武汉大学中南医院儿科;武汉大学生命科学院;
【分类号】：R329.2

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