Ena/VASP家族对人巨核细胞白血病Dami细胞中GPIb-IX复合物表达的影响  

The Effect of Ena/VASP Family on the Expression of GPIb-IX Complex in Human Megakaryoblastic Leukemia Dami Cells

作　　者：葛鑫鑫 袁柳霞 梁展文 李徐 闫荣[1] 戴克胜[1] GE Xin-Xin;YUAN Liu-Xia;LIANG Zhan-Wen;LI Xu;YAN Rong;DAI Ke-Sheng(The First Affiliated Hospital of Soochow University,Jiangsu Institute of Hematology,National Clinical Research Center for Hematological Diseases,Key Laboratory of Thrombosis and Hemostasis of National Health Comission,Suzhou 215000,Jiangsu Province,China)

机构地区：[1]苏州大学附属第一医院,江苏省血液研究所,国家血液系统疾病临床医学研究中心,国家卫生健康委员会血栓与止血重点实验室,江苏苏州215000

出　　处：《中国实验血液学杂志》2023年第2期483-488,共6页Journal of Experimental Hematology

基　　金：国家自然科学基金(81770117,82070121);江苏省科教强卫工程-临床医学中心资助项目(YXZXA2016002);苏州大学附属第一医院博习计划自然科学基金项目(BXQN202135)。

摘　　要：目的:探索Ena/VASP家族对人巨核细胞白血病Dami细胞中GPIb-IX复合物表达的影响。方法:设计合成针对Ena/VASP家族的Enah、EVL及VASP 3个基因的干扰siRNA,通过Lipofectamine^(TM) 2000转染人巨核细胞白血病Dami细胞株,培养48 h后收取细胞,利用qRT-PCR、Western blot和流式细胞术等方法检测巨核细胞中GPIb-IX复合物的表达情况。结果:构建了Ena/VASP家族基因干扰效果较佳的Dami细胞株。在敲减Enah、EVL及VASP基因表达后,在mRNA水平上,巨核细胞中GPIb-IX复合物的表达无明显变化;在总蛋白和膜表面蛋白水平上,敲减Enah后,巨核细胞中GPIb-IX复合物表达明显降低。结论:Enah基因可能对人巨核细胞白血病Dami细胞中GPIb-IX复合物的表达有影响,但是具体机制仍需要进一步探索。Objective:To explore the effects of Ena/VASP gene family on the expression of glycoprotein(GP)Ib-IX complex in human megakaryoblastic leukemia Dami cells.Methods:SiRNAs targeting Ena/VASP gene family were designed and synthesized to interfere Enah,EVL and VASP gene expression.When the siRNAs were transfected into Dami cells by using Lipofectamine^(TM) 2000 for 48 h,the expression of GPIb-IX complex was detected by quantitative real-time PCR,Western blot and flow cytometry.Results:We successfully established siVASP,siEVL and siEnah Dami cell lines.And it was found that the expression of GPIb-IX complex had no evident reduction in siEVL or siVASP Dami cells at both mRNA and protein level,while the total protein and membrane protein of GPIb-IX complex were obviously reduced when Enah was knocked down.Conclusion:Enah could affect the expression of GPIb-IX complex in human megakaryoblastic leukemia Dami cells,but the underlying mechanism still needs to be further explored.

关 键 词：Ena/VASP家族 Dami细胞 转染 GPIb-Ⅸ复合物 

分 类 号：R733.7[医药卫生—肿瘤]