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目的研究microRNA-126(miR-126)对血管内皮细胞生长发育的影响,探寻miR-126可能参与的血管生成过程。方法常规培养脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)。构建miR-126过表达载体并转染HUVECs;实验分为:正常细胞组,空载慢病毒转染组,miR-126过表达慢病毒转染组(n=6);于转染第6天后用qRT-PCR检测内皮生长负性调节因子spred1、Pik3R2的mRNA表达水平,Western blot检测spred1、Pik3R2的蛋白表达水平。结果转染第6天后,对qRT-PCR及Western blot检测结果进行分析,过表达组中spread1、Pik3R2的2(-ΔΔCt)值较其余两组低;过表达组中spred1/GAPDH和Pik3R2/GAPDH值也均小于其余两组,差异具有统计学意义(P<0.05)。spred1、Pik3R2的mRNA表达水平明显下调,其蛋白表达水平也明显降低。结论 miR-126能促进VEGF的内皮细胞增殖及血管生成作用。
Objective To investigate the effect of microRNA-126 (miR-126) on the growth and development of vascular endothelial cells and explore the possible angiogenic process of miR-126. Methods Human umbilical vein endothelial cells (HUVECs) were cultured routinely. To construct miR-126 overexpression vector and transfect it into HUVECs. The experiment was divided into normal cell group, empty lentiviral transfection group and miR-126 overexpression lentiviral transfection group (n = 6) QRT-PCR was used to detect the mRNA expression of spred1 and Pik3R2, and Western blot was used to detect the protein expression of spred1 and Pik3R2. Results After 6 days of transfection, the results of qRT-PCR and Western blot analysis showed that the 2 (-ΔΔCt) values of spread1 and Pik3R2 in the overexpression group were lower than those in the other two groups. The levels of spred1 / GAPDH and Pik3R2 / GAPDH Values were also less than the other two groups, the difference was statistically significant (P <0.05). spred1, Pik3R2 mRNA expression was significantly down-regulated, and its protein expression levels were significantly lower. Conclusion miR-126 can promote endothelial cell proliferation and angiogenesis in VEGF.