CAJ | 학술논문

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(PLK4).将含有PLK4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HEK293T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Western blot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HEK293T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PLK4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PLK4的3'UTR结合并下调其mRNA和蛋白质的表达.
목적:분석miRNA-34c재궁경암조직중적표체수평병감정기신적파기인.방법:이용형광실시정량PCR(qRT-PCR)검측감숙성부유보건의원34례궁경암화암방정상조직중miRNA-34c표체수평.이용miRNA파기인예측수거고예측miRNA-34c신적파기인Polo양격매4(PLK4).장함유PLK4적3'UTR편단형광소매재체분별여miRNA-34c모의물혹음성대조공전염HEK293T세포,병검측기형광소매활성.재인궁경암세포SiHa세포중분별전염miRNA-34c모의물혹음성대조,이용qRT-PCR법화Western blot법분별검측파기인적mRNA화단백질적표체수평.결과:여암방정상조직상비,miRNA-34c재궁경암조직중표체하조.재HEK293T세포중,miRNA-34c모의물현저억제형광소매적활성(P<0.01).miRNA-34c모의물현저하조SiHa세포중PLK4적mRNA화단백질적표체(P<0.01).결론:miRNA-34c재인궁경암조직중표체하조,차능여PLK4적3'UTR결합병하조기mRNA화단백질적표체.
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 34 paired 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 PLK4 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 Western blot, 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.