靶向daintain/AIF-1的siRNA抑制乳腺癌细胞MDA-MB-231的增殖和迁移  

Daintain/AIF-1-Targeting Sirna Inhibits the Proliferation and Migration of MDA-MB-231

作　　者：刘收[1] 陈正望[1] 

机构地区：[1]华中科技大学生命科学与技术学院分子生物物理学教育部重点实验室,武汉市430074

出　　处：《医学分子生物学杂志》2008年第3期216-220,共5页Journal of Medical Molecular Biology

基　　金：国家自然科学基金(No.30470823)~~

摘　　要：目的构建靶向daintain/AIF-1的siRNA表达载体pRNAT-H1.1-daintain/AIF-1(pRNAT-DT),研究靶向daintain/AIF-1的siRNA对乳腺癌细胞MDA-MB-231增殖和迁移的影响。方法设计合成靶向dain-tain/AIF-1的siRNA模板双链,将其克隆入siRNA空载体pRNAT-H1.1,用脂质体法导入乳腺癌细胞MDA-MB-231中;RT-PCR和Western印迹方法检测daintain/AIF-1以及cyclinD1的表达;MTT方法检测细胞增殖;流式细胞仪检测细胞周期;Trans-well细胞板检测细胞迁移。结果针对序列1和序列2的siRNA表达载体pRNAT-DT均能有效抑制daintain/AIF-1的表达。靶向daintain/AIF-1的siRNA抑制细胞增殖和cyclinD1表达,阻滞细胞周期由S期向G2/M期转变。同时,daintain/AIF-1表达的下调抑制了细胞的迁移。结论靶向daintain/AIF-1的siRNA抑制乳腺癌细胞MDA-MB-231的增殖和迁移。Objective To construct daintain/AIF-l-targeting siRNA-expressing plasmid pRNAT-DT, and study the effect of daintain/AIF-1 on the proliferation and migration of MDA-MB- 231, to provide a basis for further research on the association of daintain/AIF-1 with the development of cancers. Methods DNA fragment corresponding to siRNA targeting daintain/AIF-1 was synthesized and cloned into plasmid pRNAT-H1.1, and the recombinant plasmid was transfected into breast cancer ceils by liposome. RT-PC, R and Western blotting were used to determine the expression of daintain/AIF-1 and cyclin D1. The proliferation of MDA-MB-231 was examined by MTT incorporation. Cell cycles were determined with flow cytometry （FCM） . Cell migration was examined by using Transwell plate. Results The expression of daintain/AIF-1 was efficiently inhibited by the siRNA-expressing plasmid pRNAT-DT targeting sequence 1 and 2. The siRNA targeting daintain/ AIF-1 gene inhibited the proliferation of MDA-MB-231, arrested the cell cycle transition from S phase to G2/M phase and decreased the expression of cyclin D1. Furthermore, the down-regulation of daintain/AIF-1 expression inhibited the migration of MDA-MB-231. Conclusion Daintain/AIF- 1-targeting siRNA can inhibit the proliferation via down-regulating the expression of cyclin D1 and restrained the migration of MDA-MB-231.

关 键 词：细胞因子daintain/AIF-1 SIRNA 增殖 迁移 

分 类 号：Q813[生物学—生物工程]