發(fā)布時(shí)間：2018-06-23 05:14

  本文選題：人角膜緣干細(xì)胞 + microRNAs��； 參考：《廣東醫(yī)學(xué)》2017年08期

【摘要】：目的探討microRNA-31(miR-31)對(duì)在胚胎干細(xì)胞微環(huán)境中培養(yǎng)的角膜緣干細(xì)胞(LSCs)自我更新與分化的調(diào)控作用。方法采用CnT-20,添加20%胚胎干細(xì)胞培養(yǎng)上清液(ESC-CM),對(duì)LSCs進(jìn)行培養(yǎng)。將ESC-CM組細(xì)胞進(jìn)行轉(zhuǎn)染miR-31或者Antago-31,檢測(cè)細(xì)胞克隆形成能力的改變;流式細(xì)胞儀分析細(xì)胞周期、凋亡的改變,qPCR檢測(cè)FIH-1、P21、P63、ABCG2、CK3、miR-31、miR-143、miR-145、miR-184的表達(dá)情況;免疫熒光與Western blot檢測(cè)CK3、Cx43 P63、ABCG2、Survivin、FIH-1、P21、Caspase-3等的表達(dá)情況。結(jié)果 miR-31 mimic組細(xì)胞的CFE明顯低于miR-31 inhibitor組[(3.00±1.00)%vs.(6.90±0.79%),P0.05];Antago-31組細(xì)胞的CFE[(8.90±0.53)%vs.(7.27±0.80),P0.05]則顯著高于Ir-antago組。miR-31mimic組進(jìn)入S期細(xì)胞的比例[(9.33±2.88)%vs.(16.00±1.73)%,P0.05]明顯低于miR-31 inhibitor組;Antago-31組進(jìn)入S期細(xì)胞的比例[(22.33±2.58)%vs.(15.33±0.58)%,P0.05]則顯著高于Ir-antago組。miR-31mimic組細(xì)胞凋亡的比例[(31.13±7.56)%vs.(4.83±0.38%),P0.05]明顯高于miR-31 inhibitor組;Antago-31組細(xì)胞凋亡的比例[(3.03±0.23)%vs.(5.93±1.53%),P0.05]則顯著低于Ir-antago組。在進(jìn)行miR-31mimc處理之后,P63、ABCG2、Survivin、FIH-1、miR-143、miR-145的表達(dá)量下降,而P21、Cx-43、CK3、miR-31、miR-184的表達(dá)量明顯上升(P0.05)。在antago-31處理組,P63、ABCG2、Survivin、FIH-1、miR-143、miR-145的表達(dá)量上升,而P21、Cx-43、CK3、miR-31、miR-184的表達(dá)量明顯下降(P0.05)。Caspase-3的表達(dá)水平在antago-31處理組一直維持在相對(duì)較低的水平。結(jié)論通過轉(zhuǎn)染Antago-31提高了FIH-1在LSCs中的表達(dá)水平,并減少了P21的表達(dá)。miR-31表達(dá)水平的下調(diào)促進(jìn)了LSCs干性的維持。ESC-CM通過抑制miR-31/FIH-1/P21部分調(diào)控了LSCs的更新與分化。
[Abstract]:Objective to investigate the effect of microRNA-31 (miR-31) on the self-renewal and differentiation of limbal stem cells (LSCs) cultured in embryonic stem cell microenvironment. Methods LSCs were cultured with CnT-20 and 20% ESC-CM. ESC-CM cells were transfected with miR-31 or Antago-31to detect the change of cell clone formation ability, the cell cycle was analyzed by flow cytometry, and the apoptosis was detected by qPCR. 緇撴灉 miR-31 mimic緇勭粏鑳?yōu)鐨凜FE鏄庢樉浣庝簬miR-31 inhibitor緇刐(3.00鹵1.00)%vs.(6.90鹵0.79%),P0.05];Antago-31緇勭粏鑳?yōu)鐨凜FE[(8.90鹵0.53)%vs.(7.27鹵0.80),P0.05]鍒欐樉钁楅珮浜嶪r-antago緇，

本文編號(hào)：2055940