miR-199a-5p调控DDR1抑制人脑胶质瘤细胞增殖和迁移

目的初步探究miR-199a-5p对人脑胶质瘤细胞增殖和迁移的影响。方法选取U251细胞为实验对象,构建miR-199a-5p过表达U251细胞株。实验分组:对照组(无转染的U251细胞,Control)、阴性对照组(转染空载体质粒U251细胞,NC)以及实验组(转染miR-199a-5p成熟模拟物,mimics)。实时荧光定量PCR检测各组细胞miR-199a-5p表达;CCK-8检测转染miR-199a-5p后细胞增殖;细胞划痕实验与Transwell迁移实验检测各组U251迁移情况;Western blot检测DDR1表达;构建DDR1过表达U251细胞株,检测DDR1过表达对转染miR-199a-5p的U251细胞增殖和迁移的影响。结果mimics组细胞miR-199a-5p水平高于control组(P<0.01),细胞活力降低(P<0.01),增殖能力减弱(P<0.01)。转染miR-199a-5p组细胞DDR1表达水平降低(P<0.01)。与mimins组相比,转染pcDNA3.1-DDR1组能够上调DDR1(P<0.01),增加细胞活力,促进细胞增殖(P<0.05或P<0.01)。结论miR-199a-5p能够下调DDR1表达,抑制人脑胶质瘤细胞增殖和迁移。

miR-199a-5p regulates DDR1 to inhibit proliferation and migration of human glioma cells

Aim To investigate the effect of miR-199a-5p on the proliferation and migration of human glioma cells.Methods U251 cells were selected as experimental subjects to construct a U251 cell line overexpressing miR-199a-5p.The experiment was divided into:control group(U251 cells without transfection,Control),negative control group(transfected with empty vector plasmid U251 cells,NC)and experimental group(transfected with miR-199a-5p mature mimics,mimics).Real-time fluorescent quantitative PCR was used to detect the expression of miR-199a-5p in each group;CCK-8 was used to detect the proliferation of cells transfected with miR-199a-5p;the cell scratch test and Transwell migration test were used to detect the migration of U251 in each group;Western blot was applied to detect DDR1 expression;a U251 cell line overexpressing DDR1 was constructed to detect the effect of overexpression of DDR1 on the proliferation and migration of U251 cells transfected with miR-199a-5p.Results The level of miR-199a-5p in mimics group was significantly higher than that in control group(P<0.01),the cell viability was reduced(P<0.01),and the proliferation ability was weakened(P<0.01).The expression of DDR1 in miR-199a-5p group cells was significantly reduced(P<0.01).Compared with mimincs group,the pcDNA3.1-DDR1 transfected group could up-regulate DDR1(P<0.01),increase cell viability,and promote cell proliferation(P<0.05 or P<0.01).Conclusions miR-199a-5p can down-regulate the expression of DDR1 and inhibit the proliferation and migration of human glioma cells.