【摘要】：目的探討miR-24對(duì)人臍靜脈內(nèi)皮細(xì)胞(HUVECs)增殖、轉(zhuǎn)移、自噬的影響。方法將HUVECs隨機(jī)分為空白對(duì)照組、雷帕霉素+miR-24高表達(dá)組、雷帕霉素組。空白對(duì)照組不給予任何處理;雷帕霉素組的HUVECs用1 000 nmol/L雷帕霉素處理6 h建立自噬模型。雷帕霉素+miR-24高表達(dá)組的HUVECs先轉(zhuǎn)染miR-24高表達(dá)質(zhì)粒,待轉(zhuǎn)染成功后以同樣方法建立自噬模型;用CCK-8法檢測(cè)HUVECs增殖能力,細(xì)胞劃痕試驗(yàn)檢驗(yàn)細(xì)胞的轉(zhuǎn)移能力,進(jìn)一步用免疫組織化學(xué)和Western blotting法檢測(cè)HUVECs自噬蛋白微管相關(guān)蛋白1輕鏈3(LC3)和Beclin-1的蛋白表達(dá)水平,采用透射電鏡觀察細(xì)胞內(nèi)部自噬小體。結(jié)果與空白對(duì)照組相比,雷帕霉素組、雷帕霉素+miR-24高表達(dá)組OD值、細(xì)胞遷移比例降低、Beclin-1蛋白及LC3Ⅱ蛋白表達(dá)量上升(P0.01或0.05)。與雷帕霉素組相比,雷帕霉素+miR-24高表達(dá)組OD值、細(xì)胞遷移比例下降,Beclin-1、LC3Ⅱ蛋白表達(dá)量下降(P0.01或0.05)。透射電鏡觀察結(jié)果顯示,空白對(duì)照組和雷帕霉素+miR-24高表達(dá)組的細(xì)胞質(zhì)內(nèi)各細(xì)胞器分布基本正常,細(xì)胞核形態(tài)也正常,細(xì)胞質(zhì)中未見(jiàn)明顯自噬小體;雷帕霉素組細(xì)胞質(zhì)中可見(jiàn)明顯的空泡狀結(jié)構(gòu)、包含部分雙層膜結(jié)構(gòu)的自噬小體。結(jié)論 miR-24可顯著抑制HUVECs的增殖、轉(zhuǎn)移及自噬。
[Abstract]:Objective to investigate the effects of miR-24 on proliferation, metastasis and autophagy of human umbilical vein endothelial cells (HUVECs). Methods HUVECs was randomly divided into blank control group, rapamycin miR-24 overexpression group and rapamycin group. The HUVECs of rapamycin group was treated with 1 000 nmol/L rapamycin for 6 h to establish autophagy model. The HUVECs of rapamycin miR-24 high expression group was first transfected into miR-24 high expression plasmid, and the autophagy model was established by the same method after the transfection was successful. The proliferation ability of HUVECs was detected by CCK-8 method, the ability of cell metastasis was tested by cell scratch test, and the expression of HUVECs autophagy protein microtubule-associated protein 1 light chain 3 (LC3) and Beclin-1 was detected by immunohistochemistry and Western blotting method. The intracellular autophagy was observed by transmission electron microscopy (TEM). Results compared with the blank control group, the OD value, cell migration ratio and the expression of Beclin-1 protein and LC3鈪，

本文編號(hào)：2439257