山东医药2018，Vol.58Issue(5)：5-8,4.DOI:10.3969/j.issn.1002-266X.2018.05.002
KDM2B过表达慢病毒载体构建及其在人卵巢癌细胞中的表达
Construction of lentiviral vector with KDM2B over-expression and its expression in ovarian cancer cells
摘要
Abstract
Objective To construct the lentiviral vector with KDM2B over-expression and to explore whether it can be stably expressed in the human ovarian cancer A2780 and SKOV3 cells.Methods According to the KDM2B gene nucleotide sequence provided by Gen-Bank database,the linearized vector was obtained using restriction endonuclease,and target gene fragment was prepared and packaged with GV lentiviral vector.The positive clone recombinant products were sequenced and identified.A2780 and SKOV3 cells were infected with LV-KDM2B over-expression lentivirus (LV-KDM2B infection group),while cells infected with LV-CON were taken as the negative controls (LV-CON infection group),and the uninfected A2780 and SKOV3 cells served as the blank control group.KDM2B mRNA and protein expression levels were analyzed by real-time quantitative PCR (RT-qPCR) and Western blotting,respectively.Results The lentivirus positive clone sequencing was consistent with the target sequence.The mRNA and protein expression levels of KDM2B in the A2780 and SKOV3 cells of the LV-KDM2B group were higher than those of the LV-CON infection group and blank control group,and the differences were statistically significant (all P ＜ 0.05).Conclusion The lentiviral vector with LV-KDM2B over-expression is successfully constructed,and KDM2B gene can be stably expressed in the human ovarian cancer A2780 and SKOV3 cells.关键词
卵巢癌/赖氨酸特脱甲基酶2B/A2780细胞/SKOV3细胞Key words
ovarian carcinoma/lysine-specific demethylase 2B (KDM2B)/A2780 cells/SKOV3 cells分类
医药卫生引用本文复制引用
蒙家华,况燕,卢芳芳,易叶叶,徐红..KDM2B过表达慢病毒载体构建及其在人卵巢癌细胞中的表达[J].山东医药,2018,58(5):5-8,4.基金项目
国家自然科学基金资助项目(81360389) （81360389）
广西自然科学基金资助项目(2015GXNSFAA139152). （2015GXNSFAA139152）
