基础医学与临床2013，Vol.33Issue(2)：149-155,7.
siRNA干扰人BMP9重组腺病毒载体的构建及其促进乳腺癌SK-BR-3细胞增殖
Construction of recombinant adenovirus vector interfering the expression of human BMP9 and promoting the proliferation of breast cancer SK-BR-3 cells
摘要
Abstract
Objective To screen specific small interfering RNA(siRNA) target human BMP9 gene and to prepare recombinant adenovirus vector AdsiBMP9 for investigation of its effects on the proliferation of breast cancer SK-BR-3 cells. Methods Three pairs of double-stranded DNA fragments for silencing human BMP9 were designed and synthesized, then subcloned into the shuttle plasmid Pses-Hus. The recombinant plasmids Pses-Hus-siBMP9 were transfected into the breast epithelial cells HBL-100 by lipofectamine transfection reagent, screened the effective interfering plasmid, constructed AdsiBMP9 and infected SK-BR-3 cells. The expression level of BMP9 mRNA and protein were detected by RT-PCR and western blot. The proliferation of SK-BR-3 cells were observed with MTT assay. Results The recombinant plasmid Pses-Hus-siBMP9 and recombinant adenovirus AdsiBMP9 were successfully constructed and its titer was 1 × 1010 IU/mL. Compared to the negative and non-infected controls, the expres- sion of BMP9 gene was significantly inhibited after the SK-BR-3 cells were infected by AdsiBMP9. SK-BR-3 cells infected with AdsiBMP9 showed a promotive effect. On the fifth day, the growth rate of experiment groups was significantly higher than that of negative groups. Conclusions Specific siRNA targeting human BMP9 gene was successfully constructed, which can effectively inhibit endogenous expression of BMP9 in SK-BR-3 cells and promote its proliferation.关键词
siRNA/人BMP9/重组腺病毒载体/肿瘤增殖Key words
siRNA/ human BMP9/ recombinant adenovirus vector/ tumor proliferation分类
医药卫生引用本文复制引用
刘月红,王科,孙笑笑,万绍恒,王维,陈莹莹,张彦..siRNA干扰人BMP9重组腺病毒载体的构建及其促进乳腺癌SK-BR-3细胞增殖[J].基础医学与临床,2013,33(2):149-155,7.基金项目
国家自然科学基金(81172017 （）
30800658) （）
