CD226 is involved in megakaryocyte activation and early-stage differentiation
CD226 is involved in megakaryocyte activation and early-stage differentiation
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CD226 参与巨核细胞活化和早期分化
DOI：
10.1016/j.molimm.2019.01.013
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发表时间：
2019-03-01
影响因子：
3.6
通讯作者：
Zhang, Yuan
中科院分区：
文献类型：
--
作者：
Zhang, Jinxue;Zhuang, Ran;Zhang, Yuan
本研究旨在探讨CD 226对巨核细胞分化、活化和多倍化的影响及其可能机制。用佛波醇12-肉豆蔻酸酯13-乙酸酯诱导Dami细胞成熟。通过用CD 226特异性shRNA慢病毒载体感染使CD 226沉默。qRT-PCR检测CD 226 mRNA水平。流式细胞术检测Dami细胞表面CD 226、MK特异性标志物CD 41和CD 62 P的表达及Dami细胞和CD 226敲低（KD）细胞的DNA倍体。Western blot和共聚焦分析检测CD 226对巨核细胞相关转录因子表达的影响。转染CD 226 shRNA慢病毒后，CD 226和CD 62的表达均明显降低。CD 226基因沉默导致低倍体MK细胞形态学改变和分化迟缓。此外，CD 226敲除（KO）小鼠表现出增加的2N-4 N低倍性MK和减少的>= 8 N多倍性。有趣的是，巨核细胞中CD 226的沉默下调了早期转录因子的表达，包括GATA结合因子1（加塔-1）和朋友白血病整合因子1（FLI-1），但不下调晚期核因子红细胞2（NF-E2）。CD 226参与MKs的激活和细胞周期的多倍性调控。
This study was conducted to investigate the effect of CD226 on the differentiation, activation, and polyploidization of megakaryocytes (MKs) and explore the potential mechanism. Dami (megakaryocyte line) cell maturation was induced by phorbol 12-myristate 13-acetate. CD226 was silenced by infection with a CD226-specific shRNA lentiviral vector. The mRNA level of CD226 was detected by qRT-PCR. The expressions of Dami cells surface CD226, MK specific markers CD41 and CD62P, and DNA ploidy in Dami cells and CD226 knockdown (KD) cells were evaluated by flow cytometry. The effect of CD226 on the expression of megakaryocyte-associated transcription factors was measured by western blot and confocal analysis. Transfection with CD226 shRNA lentivirus dramatically decreased the level of CD226 and expression of CD62 Pin Dami cells. Silencing of CD226 caused morphological changes and differentiation retardation in low-ploidy MK. Furthermore, CD226 knockout (KO) mice exhibited increased 2N-4N low-ploidy MK and decreased >= 8N polyploidy. Interestingly, silencing of CD226 in megakaryocytic cells down-regulated the expression of early stage transcription factors includes GATA-binding factor 1 (GATA-1) and friend leukemia integration 1 (FLI-1), but not late-stage nuclear factor, erythroid 2 (NF-E2). CD226 is involved in MKs activation and polyploidy cell cycle control.