RNA干扰CXCR4表达抑制乳腺癌MDA-MB-231细胞的增殖、黏附和迁移  

Interfering CXCR4 expression inhibits proliferation,adhesion and migration of breast cancer MDA-MB-231 cells

作　　者：刘磊[1] 郭善禹[1] 顾岩[1] 钟强[1] 

机构地区：[1]上海交通大学医学院附属第九人民医院普外科,上海200011

出　　处：《中国肿瘤生物治疗杂志》2010年第4期408-413,共6页Chinese Journal of Cancer Biotherapy

基　　金：上海教育委员会科研基金资助(No.09yz79)~~

摘　　要：目的:构建CXC趋化因子受体4(CXC chemokine receptor4,CXCR4)RNA干扰真核表达载体,研究其对人乳腺癌细胞MDA-MB-231增殖、黏附及迁移能力的抑制作用。方法:构建针对CXCR4的带发夹结构的小RNA干扰序列,并连接到pGCsi-U6-Neo-GFP载体中,转染293T细胞,筛选出干扰效率最高的表达载体。脂质体法转染MDA-MB-231细胞。利用CCK8法、细胞-基质黏附实验和划痕修复实验检测shRNA干扰CXCR4表达对MDA-MB-231细胞增殖、黏附和迁移能力的影响。结果:成功构建CXCR4-shRNA重组质粒,并转染293T细胞,利用RT-PCR及Western blotting检测发现CXCR4沉默效率最高可达81.3%。CXCR4-shRNA转染能显著抑制MDA-MB-231细胞的增殖(P<0.05)以及细胞与细胞外基质的黏附(P<0.05)。CXCR4-shRNA转染组MDA-MB-231细胞的迁移距离明显低于对照质粒组和空白对照组(P<0.01)。结论:CXCR4-shRNA干扰载体能特异性抑制CXCR4的表达,从而抑制乳腺癌MDA-MB-231细胞的增殖、黏附及迁移。Objective：To construct short-hairpin RNA（shRNA）eukaryotic expression vector targeting CXC chemokine receptor 4（CXCR4）,and to observe its impact on the proliferation,adhesion and migration of human breast cancer MDA-MB-231 cells.Methods：The fragments of CXCR4 shRNA were synthesized and cloned into pGCsi-U6-Neo-GFP vector.The recombinant plasmids were transfected into 293T cells and the most effective interfering vector was selected.MDA-MB-231 cells were transfected by liposome assay.The effects of silencing CXCR4 expression by shRNA on the growth,adhesion and migration of MDA-MB-231 cells were determined by CCK8,cell-matrix adhesion and wound healing assays,respectively.Results：The shRNA eukaryotic expression vectors targeting CXCR4（CXCR4-shRNA）were successfully constructed and transfected into 293T cells.RT-PCR and Western blotting results showed that the maximum inhibitory rate of CXCR4 expression was 81.3%.CXCR4-shRNA transfection significantly inhibited the proliferation of MDA-MB-231 cells（P〈0.05）and the adhesion between MDA-MB-231 cells and extracellular matrix（P〈0.05）.Wound healing experiment showed that the migration distance of MDA-MB-231 cells in CXCR4-shRNA transfection group was significantly lower than those in the control plasmid and the blank control group（P〈0.01）.Conclusion：CXCR4-shRNA interfering vector can specifically inhibit CXCR4 expression,proliferation,adhesion and migration of MDA-MB-231 cells.

关 键 词：CXC趋化因子受体4 RNA干扰 真核表达载体 乳腺癌 

分 类 号：R730.54[医药卫生—肿瘤] R737.9[医药卫生—临床医学]