CPEB1对人脐静脉血管内皮细胞增殖和迁移的影响及机制

发布时间：2018-06-28 21:00

  本文选题：人脐静脉血管内皮细胞 + 胞质多聚腺苷酸结合蛋白　； 参考：《山东医药》2017年09期

【摘要】：目的探讨胞质多聚腺苷酸结合蛋白1(CPEB1)对人脐静脉血管内皮细胞(HUVECs)增殖和迁移的影响及可能机制。方法取对数生长期的HUVECs细胞,分为实验组与对照组,分别用肿瘤细胞上清液培养基和普通培养基重悬细胞,Real-time PCR法和Western blotting法分别检测CPEB1及整合素-金属蛋白酶17(ADAM17)的mRNA和蛋白表达;取对数生长期HUVECs细胞,待培养至细胞达70%~80%融合时用LipofectamineTM2000进行转染,细胞分为携带基因的质粒转染组(转染组)、不带任何基因的空质粒转染细胞组(转染对照组)和不经任何处理的细胞组(空白对照组)。MTT法检测转染后HUVECs的增殖能力,Transwell小室法检测转染后HUVECs的迁移能力,RT-PCR法检测ADAM17的mRNA表达,Western blotting法检测ADAM17蛋白表达。结果实验组中CPEB1的mRNA及蛋白表达均低于对照组(P均0.05),ADAM17的mRNA及蛋白表达均高于对照组(P均0.05);转染48、72 h后,转染组吸光度值低于转染对照组和空白对照组(P均0.05);转染组细胞穿过小室膜数目低于转染对照组和空白对照组(P均0.05);转染组ADAMl7的mRNA和蛋白表达低于转染对照组和空白对照组(P均0.05)。结论 CPEB1可抑制HUVECs细胞增殖和迁移,其机制可能与调控ADAM17表达有关。
[Abstract]:Objective to investigate the effect of cytosolic polyadenylate binding protein-1 (CPEB1) on proliferation and migration of human umbilical vein endothelial cells (HUVECs) and its possible mechanism. Methods HUVECs cells in logarithmic growth stage were divided into experimental group and control group. The mRNA and protein expression of CPEB1 and integrin-metalloproteinase-17 (ADAM17) were detected by real-time PCR and Western blotting in tumor cell supernatant medium and ordinary culture medium respectively. HUVECs cells in logarithmic growth phase were transfected with Lipofectamine TM2000 when the cells reached 70% fusion. Cells were divided into plasmid transfection group carrying gene (transfection group), empty plasmid transfection group without any gene (transfection control group) and cell group without any treatment (blank control group). MTT assay was used to detect the proliferation of HUVECs after transfection. MRNA expression of ADAM17 was detected by RT-PCR and the expression of ADAM17 protein was detected by Western blotting. Results the expression of CPEB1 mRNA and protein in the experimental group was lower than that in the control group (P 0.05). The absorbance value of transfection group was lower than that of transfection control group and blank control group (P 0.05), the number of cells passing through ventricular membrane in transfection group was lower than that in transfection control group and blank control group (P 0.05), the mRNA and protein expression of ADAMl7 in transfection group was lower than that in transfection control group. Blank control group (P 0.05). Conclusion CPEB1 can inhibit the proliferation and migration of HUVECs cells, and its mechanism may be related to the regulation of ADAM17 expression.
【作者单位】： 华中科技大学同济医学院附属武汉中心医院;华中科技大学同济医学院附属同济医院;
【基金】：华中科技大学同济医学院附属武汉中心医院院内科研基金项目(YQ14A03)
【分类号】：R730.2
，

本文编号：2079336