文章摘要
潘梅,魏钦俊,朱义保,曹 芳,曹 新.PRLR基因反义RNA对人乳腺癌细胞系MCF-7增殖的影响[J].南京医科大学学报,2007,(5):452~456
PRLR基因反义RNA对人乳腺癌细胞系MCF-7增殖的影响
The effect of PRLR antisense RNA on proliferation of human breast cancer cell MCF-7
投稿时间:2006-11-05  
DOI:10.7655
中文关键词: 人催乳素受体  反义RNA  乳腺癌  MCF-7
英文关键词: PRLR  antisense RNA  breast cancer  MCF-7
基金项目:南京医科大学创新基金(CX2002003)
作者单位
潘梅 南京医科大学生物技术系江苏 南京 210029 
魏钦俊  
朱义保  
曹 芳  
曹 新  
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中文摘要:
      目的:构建表达人催乳素受体基因(prolactin receptor,PRLR)反义RNA的真核表达质粒,观察其对乳腺癌细胞MCF-7增殖的影响。方法:应用基因重组技术,将人PRLR的cDNA反向克隆至pcDAN3.1(-)载体中,构建PRLR反义RNA真核表达质粒,将其转染入人乳腺癌细胞系MCF-7中,经G418筛选得到稳定细胞克隆,利用荧光定量PCR检测转染后细胞PRLR基因表达量的变化,应用MTT法、平板克隆形成实验及流式细胞术评价转染后细胞的增殖情况。结果:成功构建出能够表达PRLR反义RNA的真核表达质粒,转染该质粒后,MCF-7中PRLR基因表达量下降,细胞生长变慢,克隆形成能力降低,G1期细胞增加,S期细胞减少。结论:PRLR反义RNA能够下调该基因的表达,并且抑制乳腺癌细胞的增殖,证明反义基因干预治疗的可能性,为乳腺癌基因治疗的进一步研究提供了必要的依据。
英文摘要:
      Objective:To construct eukaryotic expression vector carrying anti-sense PRLR gene and investigate whether the reco-mbined plasmids could regulate the expression of PRLR mRNA and affect the growth of human breast cancer cell line MCF-7. Methods:PRLR cDNA was inserted into the expression plasmid pcDNA3.1(-) to construct the recombined plasmids p-anti-PRLR that encoding PRLR cDNA in an anti-sense orientation, then the recombined plasmids transfected MCF-7 cells by lipofectin. The stable cell lines were obtained after G418 selection. PRLR mRNA expression was detected by real-time PCR. MTT assay, clone formation test, and the flow cytometry(FCM) were used to evaluate cell proliferation. Results:(1)The eukaryotic expression vector p-anti-PRLR was constructed successfully. (2)Antisense PRLR gene decreased the level of PRLR mRNA in MCF-7 cells. (3)After antisense gene transfection, the proliferation ability of MCF-7 cell was decreased, and the G1 phase cells were increased, but the S phase cells were decreased in cell cycle. Conclusion:The results suggested that antisense PRLR could inhibit MCF-7 cell proliferation, and it might be significant for the breast cancer gene therapy.
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