siRNA沉默EPAS1基因对低氧条件下大鼠PASMCs增殖的影响
发布时间：2018-10-24 16:15
【摘要】：目的探讨应用siRNA技术沉默EPAS1(HIF-2α)基因表达对低氧条件下培养的大鼠肺动脉平滑肌细胞(PASMCs)增殖的影响。方法原代培养大鼠PASMCs共采用免疫荧光法进行鉴定。构建特异性EPAS1 siRNA脂质体,转染PASMCs,从3个干扰靶点中选取效果最好的靶点进行干扰;在常氧(37℃、5%CO_2、20%O_2)和低氧(37℃、5%CO_2、2%O_2)条件下分别培养24、48、72h,采用Western blotting检测PASMCs中EPAS1、VEGF蛋白的表达水平,CCK-8法检测细胞增殖情况。结果成功分离培养原代大鼠PASMCs并经免疫荧光鉴定证实。将特异性的EPAS1 siRNA脂质体转染到PASMCs,选择干扰效果最好的靶点2进行干扰。与常氧条件比较,低氧条件下PASMCs中VEGF蛋白的表达及细胞增殖水平均明显升高;沉默EPAS1后,无论低氧还是常氧条件下,PASMCs中VEGF蛋白的表达及细胞增殖水平均明显降低。结论 EPAS1基因参与了低氧条件下大鼠PASMCs增殖的调控,其调节可能是通过VEGF介导完成的。
[Abstract]:Objective to investigate the effect of silencing EPAS1 (HIF-2 伪) gene expression by siRNA technique on the proliferation of rat pulmonary artery smooth muscle cells (PASMC) cultured in hypoxia. Methods the primary cultured rat PASMCs was identified by immunofluorescence assay. The specific EPAS1 siRNA liposomes were constructed and transfected into PASMCs, to select the most effective target for interference. The expression level of EPAS1,VEGF protein in PASMCs was detected by Western blotting and the proliferation of cells was detected by CCK-8 method in normal oxygen (37 鈩，
本文编号：2291881
[Abstract]:Objective to investigate the effect of silencing EPAS1 (HIF-2 伪) gene expression by siRNA technique on the proliferation of rat pulmonary artery smooth muscle cells (PASMC) cultured in hypoxia. Methods the primary cultured rat PASMCs was identified by immunofluorescence assay. The specific EPAS1 siRNA liposomes were constructed and transfected into PASMCs, to select the most effective target for interference. The expression level of EPAS1,VEGF protein in PASMCs was detected by Western blotting and the proliferation of cells was detected by CCK-8 method in normal oxygen (37 鈩，
本文编号：2291881
本文链接：https://www.wllwen.com/kejilunwen/jiyingongcheng/2291881.html
最近更新
教材专著
