本文關(guān)鍵詞：微小RNA-223-3p通過調(diào)節(jié)MYH10基因促進(jìn)巨核細(xì)胞多倍體化　出處：《軍事醫(yī)學(xué)》2017年07期 　論文類型：期刊論文

  更多相關(guān)文章： miR--p MYH 巨核細(xì)胞 聚合酶鏈反應(yīng) 流式細(xì)胞術(shù)

【摘要】：目的探討微小RNA-223-3p(miR-223-3p)對巨核細(xì)胞分化和成熟的影響,并初步探索其中可能的機(jī)制。方法通過實時定量PCR檢測巨核細(xì)胞分化過程中miR-223-3p的內(nèi)源性表達(dá)變化趨勢,而后外源調(diào)節(jié)miR-223-3p在細(xì)胞系中的表達(dá)量,并通過流式細(xì)胞術(shù)檢測其對于巨核細(xì)胞分化和成熟的影響,運用生物信息學(xué)分析,找到其發(fā)揮相關(guān)生物學(xué)作用的靶基因MYH10,并通過實時定量PCR、熒光素酶、流式細(xì)胞術(shù)驗證MYH10是miR-223-3p的靶基因。結(jié)果內(nèi)源性miR-223-3p隨著巨核細(xì)胞的分化成熟表達(dá)量增加,在K562和Meg-01細(xì)胞系中轉(zhuǎn)染miR-223-3p mimics后可升高巨核細(xì)胞相關(guān)表面標(biāo)志CD41和CD61的陽性率,同時顯著促進(jìn)多倍體的形成,MYH10的基因表達(dá)隨miR-223-3p表達(dá)升高而下降,通過雙熒光素酶報告基因技術(shù)驗證MYH10基因是miR-223-3p的靶基因,進(jìn)一步抑制MYH10基因表達(dá)后,可促進(jìn)巨核細(xì)胞多倍體化。結(jié)論 miR-223-3p可通過調(diào)控MYH10的表達(dá)調(diào)節(jié)巨核細(xì)胞的成熟。
[Abstract]:Objective to investigate the effects of microRNA-223-3pmmiR-223-3p) on the differentiation and maturation of megakaryocytes. Methods the endogenous expression of miR-223-3p in megakaryocyte differentiation was detected by real-time quantitative PCR. Then exogenous regulation of miR-223-3p expression in the cell line, and flow cytometry to detect its effect on megakaryocyte differentiation and maturation, bioinformatics analysis. The target gene MYH10 was found to play a related biological role, and real-time quantification of PCR-, luciferase. Flow cytometry confirmed that MYH10 was the target gene of miR-223-3p. Results the expression of endogenous miR-223-3p increased with the differentiation and maturation of megakaryocytes. Transfection of miR-223-3p mimics into K562 and Meg-01 cells increased the positive rate of CD41 and CD61. At the same time, the gene expression of MYH10 decreased with the increase of miR-223-3p expression. Double luciferase reporter gene technique was used to confirm that MYH10 gene is the target gene of miR-223-3p, and further inhibit the expression of MYH10 gene. Conclusion miR-223-3p can regulate the maturation of megakaryocyte by regulating the expression of MYH10.
【作者單位】： 南方醫(yī)科大學(xué);軍事醫(yī)學(xué)科學(xué)院野戰(zhàn)輸血研究所干細(xì)胞與再生醫(yī)學(xué)研究室;華南生物醫(yī)藥研究院;
【基金】：國家863計劃資助項目(2013AA020107) 國家自然科學(xué)基金資助項目(81301132) 北京市自然科學(xué)基金資助項目(7152108) 廣州市健康醫(yī)療協(xié)同創(chuàng)新重大專項資助項目(201508020257,201400000003-4)
【分類號】：R3416
【正文快照】： 血小板是由骨髓中巨核細(xì)胞生成的具有凝血止血功能的小塊胞質(zhì),其在人體外周血中的數(shù)量如50×109/L,會導(dǎo)致機(jī)體出血傾向,當(dāng)血小板嚴(yán)重減少時,將威脅到人體的生命。目前臨床上主要采用機(jī)采血小板輸注來維持血小板數(shù)量,而機(jī)采血小板依賴于獻(xiàn)血者,且由于獻(xiàn)血時間長,舒適度低,血小，

本文編號：1372963