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目的分析喉癌中S100A8基因3’UTR与其靶向miRNA的调控作用。方法构建S100A8基因3′UTR野生型和突变型荧光素酶载体,应用MiRanda和RNA22软件预测S100A8基因3’UTR靶向miRNA,共转构建的荧光素酶载体和靶miRNA前体于喉癌Hep2细胞中,双荧光素酶检测系统测定荧光素酶活性。结果成功构建了S100A8基因3’UTR荧光素酶载体,在S100A83’UTR第3bp位置预测到一个miR-24结合位点,与转染controlmiR组和突变型载体组相比,miR-24前体能明显降低野生型载体的荧光素酶活性(P<0.05)。结论 miR-24靶向负性调控S100A8基因3’UTR活性。
Objective To analyze the regulatory role of 3’UTR and its targeted miRNA in S100A8 gene in laryngeal cancer. Methods The 3’UTR wild-type and mutant-type luciferase vectors of S100A8 gene were constructed. MiRanda and RNA22 software were used to predict the 3’UTR-targeted miRNA of S100A8 gene. The co-transfection of luciferase vector and target miRNA precursor in Hep2 cells In the dual luciferase assay system, luciferase activity was measured. Results The 3’UTR luciferase vector of S100A8 gene was successfully constructed, and a miR-24 binding site was predicted at the 3bp position of S100A83’UTR. Compared with the controlmiR group and the mutant vector group, the miR-24 precursor was significantly Reduce the luciferase activity of wild-type vector (P <0.05). Conclusion miR-24 targeting negatively regulates 3’UTR activity of S100A8 gene.