CPEB1对人脐静脉血管内皮细胞增殖和迁移的影响及机制

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ADAMU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.%Objective To investigate the effect of cytoplasmic polyadenylation-element-binding protein 1 (CPEB1) on the proliferation and migration of human umbilical-vein endothelial cells (HUVECs) and its possible mechanism.Methods HUVECs in logarithmic phase were divided into experimental group and control group,which were cultured with tumor cell supernatant medium and the common medium,respectively.Real-time PCR and Western blotting were used to detect the mRNA and protein expression of CPEB1 and a disintegrin and metalloproteinase 17 (ADAM17).HUVECs in logarithmic phase with 70％-80％ cells conjugation were transfected by LipofectamineTM2000.Cells were then divided into three group:carrying gene plasmid transfection group (transfection group),without empty plasmid gene transfection group (control group) and without any processing cell group (blauk control group).The proliferation ability of HUVECs after transfection was determined by MTT,the migration ability of HUVECs was detected by Transwell chamber,and RT-PCR was used to detect ADAM17 mRNA expression and Western blotting to detect ADAM17 protein expression.Results CPEB1 mRNA and protein expression levels in the experimental group were lower than those of the control group (all P ＜ 0.05).ADAM17 mRNA and protein the expression levels in experimental group were higher than those of the control group (all P ＜0.05).At 48 h and 72 b after transfection,the absorbance value (A) of the transfection group was obviously lower than those of control group blank control group (all P ＜ 0.05).The number of cells through the small chamber membrane in the transfection group was lower than that in the control group and blank control group (all P ＜ 0.05).ADAM17 mRNA and protein levels in the transfection group was lower than that in the control group and blank control group (all P ＜ 0.05).Contusion The supematant of tumor cells can induce the change in expression of CPEB1 and ADAM17 in HUVECs,and over-expression of CPEB1 may inhibit the proliferation and migration of HUVECs by regulating ADAM17.