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MicroRNAs(miRNAs)是一类大约22个核苷酸的非编码RNA.它能通过调控生长因子表达,引发肿瘤形成、细胞增殖和组织器官发育.本文通过构建miR-129-5p靶基因序列的双荧光素酶报告载体分析了miR-129-5p与靶基因之间的关系,应用脂质体转染技术和实时荧光定量技术观察了miR-129-5p在小鼠乳腺上皮细胞中的表达量及其变化,通过CASY细胞活力仪检测转染后的细胞增殖与活力变化,采用Western印迹方法检Igf-1的变化.结果表明:miR-129-5p在小鼠乳腺青春期表达最高,成功构建了Igf-1基因3’UTR荧光素酶报告载体,miR-129-5p抑制其荧光素酶活性(P<0.01),转染抑制子后miR-129-5p表达降低(P<0.01),胰岛素样生长因子(Igf-1)表达增强(P<0.05),细胞增殖和活力增强(P<0.01),结果提示:miR-129-5p可能通过抑制靶基因蛋白Igf-1的表达,进而抑制小鼠乳腺上皮细胞增殖和活力.
MicroRNAs (miRNAs) are a class of non-coding RNA of about 22 nucleotides which can induce tumor formation, cell proliferation and tissue and organ development by regulating the expression of growth factors.In this paper, The luciferase reporter vector was used to analyze the relationship between miR-129-5p and target genes. The expression of miR-129-5p in mouse mammary epithelial cells was detected by lipofection and real-time fluorescence quantitative analysis. The changes of cell proliferation and viability were detected by CASY cell viability assay and the changes of Igf-1 were detected by Western blotting.The results showed that miR-129-5p was the highest expression in mouse mammary gland during adolescence, and Igf MiR-129-5p inhibited the luciferase activity (P <0.01), the expression of miR-129-5p decreased (P <0.01), the insulin-like growth The results showed that miR-129-5p could inhibit the expression of Igf-1, and then inhibit the expression of Igf-1 in mouse mammary gland (P <0.05) Epithelial cell proliferation and vitality.