miR-24对人脐静脉内皮细胞增殖、转移及自噬的影响
发布时间：2019-03-13 09:08
【摘要】：目的探讨miR-24对人脐静脉内皮细胞(HUVECs)增殖、转移、自噬的影响。方法将HUVECs随机分为空白对照组、雷帕霉素+miR-24高表达组、雷帕霉素组。空白对照组不给予任何处理;雷帕霉素组的HUVECs用1 000 nmol/L雷帕霉素处理6 h建立自噬模型。雷帕霉素+miR-24高表达组的HUVECs先转染miR-24高表达质粒,待转染成功后以同样方法建立自噬模型;用CCK-8法检测HUVECs增殖能力,细胞划痕试验检验细胞的转移能力,进一步用免疫组织化学和Western blotting法检测HUVECs自噬蛋白微管相关蛋白1轻链3(LC3)和Beclin-1的蛋白表达水平,采用透射电镜观察细胞内部自噬小体。结果与空白对照组相比,雷帕霉素组、雷帕霉素+miR-24高表达组OD值、细胞迁移比例降低、Beclin-1蛋白及LC3Ⅱ蛋白表达量上升(P0.01或0.05)。与雷帕霉素组相比,雷帕霉素+miR-24高表达组OD值、细胞迁移比例下降,Beclin-1、LC3Ⅱ蛋白表达量下降(P0.01或0.05)。透射电镜观察结果显示,空白对照组和雷帕霉素+miR-24高表达组的细胞质内各细胞器分布基本正常,细胞核形态也正常,细胞质中未见明显自噬小体;雷帕霉素组细胞质中可见明显的空泡状结构、包含部分双层膜结构的自噬小体。结论 miR-24可显著抑制HUVECs的增殖、转移及自噬。
[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，
本文编号：2439257
[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，
本文编号：2439257
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