基础医学与临床 ›› 2009, Vol. 29 ›› Issue (12): 1286-1290.

• 研究论文 • 上一篇    下一篇

CXCR4-siRNA表达载体的构建及其对乳腺癌细胞侵袭能力的影响

冯俊飞 董坚 洪敏 高嫦娥   

  1. 昆明医学院第一附属医院生物治疗中心 昆明医学院第一附属医院
  • 收稿日期:2008-12-15 修回日期:2009-05-04 出版日期:2009-12-20 发布日期:2009-12-20
  • 通讯作者: 董坚

Construction of a siRNA Expression Plasmid Targeting CXCR4 and Its Effect on the invasion ability of the breast cancer cell

Jun-fei FENG, Jian DONG, Min HONG, Chang-e GAO   

  1. Biotherapy Research Center,the First Affiliated Hospital of Kunming Medical College the 1st Affiliated Hospital of Kunming Medical College
  • Received:2008-12-15 Revised:2009-05-04 Online:2009-12-20 Published:2009-12-20
  • Contact: Jian DONG,

摘要: 目的 构建趋化因子受体(CXCR4)小分子干扰RNA ( siRNA) 表达载体,研究其对乳腺癌细胞侵袭能力的影响。方法 选择CXCR4高表达的乳腺癌MDA-MB-231细胞株,设计合成人CXCR4基因不同靶点的能编码siRNA的2条双链DNA序列,克隆到真核表达载体pGE-1-U6/kna中构建siRNA表达载体,体外脂质体介导转染MDA-MB-231细胞,用Western blot分析CXCR4蛋白表达,transwell 小室检测细胞的侵袭能力。结果 成功构建了CXCR4-siRNA表达载体,瞬时转染乳腺癌细胞后能显著降低CXCR4的蛋白表达,可有效抑制人类乳腺癌MDA-MB-231细胞的侵袭能力。结论 CXCR4- siRNA表达载体通过降低CXCR4基因的蛋白表达能显著抑制人类乳腺癌细胞的侵袭能力,有望为乳腺癌转移的基因治疗开辟新途径。

关键词: 乳腺癌, CXCR4, 小干扰RNA, RNA干扰

Abstract: Objective To construct and identify the siRNA eukaryotic expression vector targeting gene CXC chemokine receptor-4 and study its role in invasion ability of bread cancer cells. Methods Two siRNAs were designed and synthesized according to the coding the cDNA sequence of CXCR4 gene and cloned into eukaryotic expression plasmid pGE-1-U6/kna.The constructed CXCR4-siRNA expression vector was transfected into MDA-MB-231 cells by liposome.Western blotting were used to evaluate the suppression of CXCR4 expression in different groups. The invasion and migration of MDA-MB-231 cells were evaluated by cell invasion assay in vitro. Results Enzyme digestion and DNA sequencing confirmed that the CXCR4-siRNA expression vector was constructed successfully. After transfection,the CXCR4-siRNA obviously suppressed the expression of CXCR4 compared with control groups and the ability of cell migration was decreased markedly. Conclusions CXCR4-siRNA expression vector can effectively suppress CXCR4 expression in the breast cancer cell and decrease potential of cell invasion, which may provide a novel applicable strategy for gene therapy of breast cancer metastasis.

Key words: breast cancer, CXCR4 gene, siRNA, RNAi

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