安徽医科大学学报2017，Vol.52Issue(8)：1099-1104,6.DOI:10.19405/j.cnki.issn1000-1492.2017.08.001
miR-205慢病毒表达载体的构建及其对乳腺癌细胞作用的初步研究
Construction of lentiviral vector of miR-205and primarily investigation of its effect on breast cancer cell
摘要
Abstract
Objective To construct a lentivirus vector of microRNA-205(miR-205), and transfect into MB231 cell line, to establish a stable cell line expressing miR-205.Methods Design and synthesis miR-205 primers, the target gene was amplified by PCR, and connect to the lentivirus expression plasmid.Packaging and testing virus drops Lv-miR-205, after the double enzyme digestion of the recombinant plasmid.Infecting MB231 cells with the Lv-miR-205,then detect the expression of miR-205 by Real Time qPCR and access the cell proliferation and migration by MTT assay and wound-healing assay.Results The DNA sequencing prove that the lentivirus vector was constructed successfully.Lv-miR-205 stablized infect MB231 cells, the expression of miR-205 elevated apparently, cell proliferation and migration was inhibited sharply.Conclusion The miR-205 lentiviral expression vector is successfully constructed and the cell line MB231 stably expressing miR-205 is established, which indicate that it could negatively regulate the malignant biological behavior of breast cancer cells, and laid the foundation for the further study of the function and mechanism of miR-205.关键词
miR-205/慢病毒/乳腺癌/细胞增殖Key words
miR-205/lentivirus/breast cancer/cell proliferation分类
医药卫生引用本文复制引用
范楚苓,张萍,孙彩虹,钱晨,戚博,赵茹,肖荣琴,刘亚坤,李菲菲..miR-205慢病毒表达载体的构建及其对乳腺癌细胞作用的初步研究[J].安徽医科大学学报,2017,52(8):1099-1104,6.基金项目
国家自然科学基金(编号:81302319) （编号:81302319）
安徽省自然科学基金(编号:1308085MH121) （编号:1308085MH121）
国家级大学生创新创业训练计划项目(编号:201510366013) （编号:201510366013）
