MKL1在巨核细胞分化中的表达及其作用的实验研究

MKL1 expression and its role in megakaryocyte differentiation

目的了解MKL1在巨核细胞分化、成熟中的作用。方法以人外周血来源CD34^+细胞巨核细胞分化为模型,采用荧光实时定量PCR法研究MKL1基因在不同分化阶段巨核细胞中的表达情况,通过构建慢病毒载体在CD34^+细胞中过表达MKL1基因,利用血细胞涂片和流式细胞仪考察MKL1过表达细胞经细胞因子刺激分化后的形态、CD41^+百分比及DNA含量。结果经qPCR研究发现,MKL1基因在成熟的巨核细胞中的表达6.8倍高于未分化的CD34^+细胞,以及2倍高于单个核巨核细胞。MKL1过表达组经诱导分化后CD41^+巨核细胞百分比与多倍体细胞百分比分别为61.5%和52.9%,均显著高于对照组36.3%和33.4%的比例。结论MKL1基因在巨核细胞分化中表达逐渐上调,其外源表达促进人外周血来源CD34^+细胞巨核细胞分化和多倍体化。

To understand the role of MKL1 in megakaryocyte differentiation and maturation.We set megakaryocyte differentiation of mobilized human peripheral blood CD34^＋ cells as studying model.Real-time quantitative PCR was used to investigate the relative MKL1 gene expression;lentivirus over-expressing MKL1 gene was constructed and used to infect PB CD34^＋ cell.Cytospin and flow cytometry was used to study the morphology,CD41^＋ expression,and DNA content.In mature megakaryocytes,the MKL1 gene expression was 6.8-fold higher than that in PB CD34^＋ cells,meanwhile more than 2-fold higher than that in mononuclear megakaryocytes.The over-expression of MKL1 in PB CD34^＋ cells promoted the differentiation and maturation of megakaryocytes in terms of CD41^＋ cells and polyploidy cells（61.5% and 52.9%,contrasting to 36.3%and 33.4%in control group）.We come to the conclusion that MKL1 gene expression is upregulated during the differentiation and its enhanced ectopic expression promotes the differentiation and maturation of megakaryocytes.