张红红, 刘晓丽, 许思娟. 宫颈癌组织中miRNA-34c 的表达水平分析及其靶基因PLK4 的鉴定[J]. 中国肿瘤临床, 2015, 42(22): 1077-1080. DOI: 10.3969/j.issn.1000-8179.2015.22.028
引用本文: 张红红, 刘晓丽, 许思娟. 宫颈癌组织中miRNA-34c 的表达水平分析及其靶基因PLK4 的鉴定[J]. 中国肿瘤临床, 2015, 42(22): 1077-1080. DOI: 10.3969/j.issn.1000-8179.2015.22.028
Honghong ZHANG, Xiaoli LIU, Sijuan XU. Analysis of miRNA- 34c expression in cervical cancer tissue and preliminary identification of Polo-like kinase4 as its target gene[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2015, 42(22): 1077-1080. DOI: 10.3969/j.issn.1000-8179.2015.22.028
Citation: Honghong ZHANG, Xiaoli LIU, Sijuan XU. Analysis of miRNA- 34c expression in cervical cancer tissue and preliminary identification of Polo-like kinase4 as its target gene[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2015, 42(22): 1077-1080. DOI: 10.3969/j.issn.1000-8179.2015.22.028

宫颈癌组织中miRNA-34c 的表达水平分析及其靶基因PLK4 的鉴定

Analysis of miRNA- 34c expression in cervical cancer tissue and preliminary identification of Polo-like kinase4 as its target gene

  • 摘要: 目的:分析miRNA-34c 在宫颈癌组织中的表达水平并鉴定其新的靶基因。方法:利用荧光实时定量PCR(qRT-PCR)检测甘肃省妇幼保健医院34例宫颈癌和癌旁正常组织中miRNA-34c 表达水平。利用miRNA 靶基因预测数据库预测miRNA-34c 新的靶基因Polo 样激酶4(PLK 4)。 将含有PLK 4 的3‘UTR 片段荧光素酶载体分别与miRNA-34c 模拟物或阴性对照共转染HEK 293T 细胞,并检测其荧光素酶活性。在人宫颈癌细胞SiHa 细胞中分别转染miRNA-34c 模拟物或阴性对照,利用qRT-PCR法和Westernblot法分别检测靶基因的mRNA 和蛋白质的表达水平。结果:与癌旁正常组织相比,miRNA-34c 在宫颈癌组织中表达下调。在HEK 293T 细胞中,miRNA-34c 模拟物显著抑制荧光素酶的活性(P < 0.01)。 miRNA-34c 模拟物显著下调SiHa 细胞中PLK 4 的mRNA 和蛋白质的表达(P < 0.01)。 结论:miRNA-34c 在人宫颈癌组织中表达下调,且能与PLK 4 的3’UTR 结合并下调其mRNA 和蛋白质的表达。

     

    Abstract: Objective:To analyze the expression profile of miRNA-34c in cervical cancer tissue and identify its novel target gene. Methods:The expression levels of miRNA-34c were detected in 34paired cervical cancer tissues and normal paraneoplastic tissues via quantitative reverse transcription polymerase chain reaction (qRT-PCR). Polo-like kinase 4 (PLK4) is a target gene of miRNA- 34c pre-dicted in the miRNA Target Database. Luciferase vector containing the binding site of miRNA-34c to PLK 4 3'UTR and miRNA- 34c mimic or negative control were co-transfected into HEK293T cells, and luciferase expression was examined. The miRNA-34c mimic or negative control was transfected into SiHa cells, and the mRNA or protein expression of PLK4 was detected via qRT-PCR or Westernblot, respectively. Results: MiRNA-34c expression was lower in cervical cancer tissues than in normal paraneoplastic tissues. The miR -NA- 34c mimics significantly inhibited luciferase activation in the HEK293T cells (P<0.01) and significantly decreased the mRNA and protein expression levels of PLK4 in the SiHa cells (P<0.01). Conclusion: MiRNA-34c is significantly decreased in cervical cancer tis-sues. Moreover, miRNA-34c can significantly repress the mRNA and protein expression levels of PLK4 by directly targeting the 3'UTR.

     

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