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

【摘要】：目的探討microRNA-31(miR-31)對在胚胎干細胞微環(huán)境中培養(yǎng)的角膜緣干細胞(LSCs)自我更新與分化的調控作用。方法采用CnT-20,添加20%胚胎干細胞培養(yǎng)上清液(ESC-CM),對LSCs進行培養(yǎng)。將ESC-CM組細胞進行轉染miR-31或者Antago-31,檢測細胞克隆形成能力的改變;流式細胞儀分析細胞周期、凋亡的改變,qPCR檢測FIH-1、P21、P63、ABCG2、CK3、miR-31、miR-143、miR-145、miR-184的表達情況;免疫熒光與Western blot檢測CK3、Cx43 P63、ABCG2、Survivin、FIH-1、P21、Caspase-3等的表達情況。結果 miR-31 mimic組細胞的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組進入S期細胞的比例[(9.33±2.88)%vs.(16.00±1.73)%,P0.05]明顯低于miR-31 inhibitor組;Antago-31組進入S期細胞的比例[(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。在進行miR-31mimc處理之后,P63、ABCG2、Survivin、FIH-1、miR-143、miR-145的表達量下降,而P21、Cx-43、CK3、miR-31、miR-184的表達量明顯上升(P0.05)。在antago-31處理組,P63、ABCG2、Survivin、FIH-1、miR-143、miR-145的表達量上升,而P21、Cx-43、CK3、miR-31、miR-184的表達量明顯下降(P0.05)。Caspase-3的表達水平在antago-31處理組一直維持在相對較低的水平。結論通過轉染Antago-31提高了FIH-1在LSCs中的表達水平,并減少了P21的表達。miR-31表達水平的下調促進了LSCs干性的維持。ESC-CM通過抑制miR-31/FIH-1/P21部分調控了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：2055940