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

作　　者：潘梅[1] 魏钦俊[1] 朱义保[1] 曹芳[1] 曹新[1] 

机构地区：[1]南京医科大学生物技术系,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2007年第5期452-456,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：南京医科大学创新基金(CX2002003)

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。Objective：To construct eukaryotic expression vector carrying anti-sense PRLR gene and investigate whether the recombined 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 panti-PRLR was constructed successfully. （2）Antisense PRLR gene decreased the level of PRLR mRNA in MCF-7 cells. （3）After antisense gene transfeetion, 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.

关 键 词：人催乳素受体 反义RNA 乳腺癌 MCF-7 

分 类 号：Q74[生物学—分子生物学] R737.9[医药卫生—肿瘤]