PD-L1蛋白过表达对HeLa细胞迁移能力的影响

发布时间：2018-03-26 04:20

本文选题：PD-L　切入点：免疫治疗　出处：《生物技术》2017年06期

【摘要】：[目的]探讨PD-L1过表达对HeLa细胞迁移的影响。[方法]通过分子克隆构建PD-L1质粒载体(p EGFPPD-L1);通过PEI法转染质粒,调节质粒转染量(1μg、3μg、5μg)和转染时间(24 h、36 h、48 h、72 h),优化转染条件;通过细胞划痕实验检测细胞融合率,Western Blot检测细胞迁移相关蛋白(E-钙黏蛋白、N-钙黏蛋白、波形蛋白)的表达量。[结果]最佳转染条件是质粒量3μg/孔,转染后48 h观察,此时转染效率可达(82.94±8.08)%。转染pEGFP-PD-L1质粒后,HeLa细胞的PD-L1过表达3.5倍,划痕融合率显著增高(p0.01);波形蛋白和N-钙黏蛋白的表达量显著升高(p0.05),E-钙黏蛋白表达量显著下降(p0.05)。[结论]PD-L1过表达显著促进HeLa细胞迁移能力和上皮向间质转化水平。
[Abstract]:[objective] to investigate the effect of overexpression of PD-L1 on the migration of HeLa cells. [methods] to construct PD-L1 plasmid vector pEGFPPD-L _ (1) by molecular cloning, to adjust the amount of 1 渭 g ~ (3 渭 g) ~ 5 渭 g ~ (5 渭 g) and the transfection time of 24 h ~ 36 h ~ 48 h ~ (72 h) to optimize the transfection conditions. The cell fusion rate and the expression of E-cadherin (vimentin) were detected by cell scratch assay and Western Blot. [results] the optimal transfection conditions were as follows: the amount of plasmid was 3 渭 g / well and 48 h after transfection, the expression of E-cadherin (vimentin) was detected. The transfection efficiency was 82.94 卤8.08. The PD-L1 expression in HeLa cells was 3.5-fold after transfection of pEGFP-PD-L1 plasmid. The expression of vimentin and N- cadherin decreased significantly. [conclusion] overexpression of PD-L1 significantly promoted the migration ability of HeLa cells and the level of epithelium to mesenchymal transformation.
【作者单位】：武汉科技大学生物医学研究院生命科学与健康学院;
【基金】：国家自然科学基金项目(“MRTF-A和STAT3调控乳腺癌EMT及机制研究”,No.31501149;“抗活性氧自由基的载白藜芦醇Ti O2纳米管促进骨修复研究”,No.31700824)
【分类号】：R737.33

【相似文献】