吉林大学学报（医学版）2016，Vol.42Issue(5)：848-854,7.DOI:10.13481/j.1671-587x.20160502
聚乙烯亚胺介导miR-2861模拟物转染MC3T3-E1细胞的体外成骨分化
Osteogenesis differentiation of MC3T3-E1 cells induced by miRNA-2861 mimic transfection mediated by polyethylenimine
摘要
Abstract
Objective:To transfect the non-viral vector polyethylenimine (PEI)mediated miR-2861 mimic into the MC3T3-E1 cell line,and to explore the transfection efficiency of PEI/miR-2861 complex and its effects on the proliferation and osteogenesis differentiation in pre-osteoblasts. Methods:The proper amount of PEI was blended with miR-2861 mimic and negative control (NC)separately in a ratio of N∶P=10∶1,and they were divided into experiment group and NC group. The NC/PEI complex acted as the NC group was used to eliminate the interference of osteogenesis from the addition of double-stranded RNA mimic.MTT assay was used to determine the optimal concentration of PEI/miR-2861 mimic complex.The fluorescence imaging technique and bulge-loop RT-PCR were used to detect the transfection efficiency and mRNA expression of miRNA-2861 in the cells with different concentrations (10,30, 50,and 100 nmol · L-1 ), separately.The osteogenesis ability of MC3T3 cells was identified with RT-PCR and Alizarin red staining with the selected concentration of PEI/miR-2861 by transient transfection.Results:Compared with blank control group,the proliferation rates of MC3T3 cells in 100 nmol·L-1 PEI/miR-2861 group was decreased significantly at 72 h (P < 0. 05 ). With the increasing of transfected concentration the transfection efficiency of miRNA/PEI complex was increased gradually.The results of Alizarin red staining and quantitative analysis showed that calcium deposits were more and bigger in experiment group after induced for 21 d,while both in blank control group and NC group they were less.Conclusion:The miRNA-2861 mimic can be effectively transfected into the MC3T3-E1 cell line and expresses with a high level,which is mediated by PEI as the gene vector.miR-2861 mimic has a certain ability of promoting osteogenesis differentiation of MC3T3-E1 cells.关键词
miRNA-2861模拟物/聚乙烯亚胺/MC3T3-E1细胞系/成骨分化/基因治疗Key words
miRNA-2861 mimic/polyethylenimine/MC3T3-E1 cells/osteogenesis differentiation/gene therapy分类
医药卫生引用本文复制引用
方滕姣子,刘杰,顾中一,宫海环,布文奂,董悦,孙宏晨..聚乙烯亚胺介导miR-2861模拟物转染MC3T3-E1细胞的体外成骨分化[J].吉林大学学报（医学版）,2016,42(5):848-854,7.基金项目
国家自然科学基金资助课题（81271111）；国家自然科学基金青年基金资助课题（81400488）；国家自然科学基金重大国际合作项目资助课题（81320108011）；吉林大学研究生创新基金资助课题 （）
