Yu Jin, Yan Feng-Juan, Han Cui-Juan, Fan Jing-Yi
Department of Pediatrics, Zhongnan Hospital of Wuhan University,Wuhan 430071, Hubei Province, China.
College of life Science, Wuhan University,Wuhan 430072, Hubei Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2017 Oct;25(5):1537-1543. doi: 10.7534/j.issn.1009-2137.2017.05.046.
To study the function of ZNF300 in the megakaryocytes differentiation and proliferation.
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.
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.
ZNF300 promotes megakaryocyte differentiation.
研究ZNF300在巨核细胞分化和增殖中的作用。
利用ZNF300表达的公共数据分析和巨核细胞培养来揭示ZNF300表达与白血病及巨核细胞分化的相关性;通过慢病毒或逆转录病毒感染介导ZNF300过表达,采用流式细胞术、MTT法和集落形成试验检测K562细胞和原代小鼠骨髓细胞向巨核细胞的分化和增殖情况;通过分离胞质和核提取物并结合蛋白质免疫印迹法检测ZNF300的亚细胞定位。采用双荧光素酶报告基因检测和染色质免疫沉淀定量PCR研究ZNF300的靶基因。
ZNF300表达上调与巨核细胞分化相关;ZNF300过表达促进CD41和CD61表达,抑制细胞周期进程,并可减少集落形成单位。ZNF300定位于细胞核并调控C-MYC表达。
ZNF300促进巨核细胞分化。