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

  更多相關文章： miR--p MYH 巨核細胞 聚合酶鏈反應 流式細胞術

【摘要】：目的探討微小RNA-223-3p(miR-223-3p)對巨核細胞分化和成熟的影響,并初步探索其中可能的機制。方法通過實時定量PCR檢測巨核細胞分化過程中miR-223-3p的內源性表達變化趨勢,而后外源調節(jié)miR-223-3p在細胞系中的表達量,并通過流式細胞術檢測其對于巨核細胞分化和成熟的影響,運用生物信息學分析,找到其發(fā)揮相關生物學作用的靶基因MYH10,并通過實時定量PCR、熒光素酶、流式細胞術驗證MYH10是miR-223-3p的靶基因。結果內源性miR-223-3p隨著巨核細胞的分化成熟表達量增加,在K562和Meg-01細胞系中轉染miR-223-3p mimics后可升高巨核細胞相關表面標志CD41和CD61的陽性率,同時顯著促進多倍體的形成,MYH10的基因表達隨miR-223-3p表達升高而下降,通過雙熒光素酶報告基因技術驗證MYH10基因是miR-223-3p的靶基因,進一步抑制MYH10基因表達后,可促進巨核細胞多倍體化。結論 miR-223-3p可通過調控MYH10的表達調節(jié)巨核細胞的成熟。
[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;軍事醫(yī)學科學院野戰(zhàn)輸血研究所干細胞與再生醫(yī)學研究室;華南生物醫(yī)藥研究院;
【基金】：國家863計劃資助項目(2013AA020107) 國家自然科學基金資助項目(81301132) 北京市自然科學基金資助項目(7152108) 廣州市健康醫(yī)療協同創(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胞生成的具有凝血止血功能的小塊胞質,其在人體外周血中的數量如50×109/L,會導致機體出血傾向,當血小板嚴重減少時,將威脅到人體的生命。目前臨床上主要采用機采血小板輸注來維持血小板數量,而機采血小板依賴于獻血者,且由于獻血時間長,舒適度低,血小，

本文編號：1372963