国际妇产科学杂志

国际妇产科学杂志 ›› 2017, Vol. 44 ›› Issue (6): 703-708.

• 论著 • 上一篇    下一篇

MicroRNA-10b对宫颈癌细胞侵袭迁移能力的影响及机制研究

陶贝贝，周文娟，张婷   

  1. 450012  郑州市妇幼保健院妇产科（陶贝贝，周文娟）；郑州大学第三附属医院妇产科（张婷）
  • 收稿日期:2017-07-18 修回日期:2017-09-27 出版日期:2017-12-15 发布日期:2017-12-15

The Effects of MicroRNA-10b on the Invasion and Migration in the Cervical Carcinoma Cells and Its Possible Mechanism

TAO Bei-bei，ZHOU Wen-juan，ZHANG Ting   

  1. Department of Obstetrics and Gynecology，Women & Infants Hospital Zhengzhou，Zhengzhou 450012，China（TAO Bei-bei，ZHOU Wen-juan）；Department of Obstetrics and Gynecology，The Third Affiliated Hospital of Zhengzhou University，Zhengzhou 450052，China（ZHANG Ting）
  • Received:2017-07-18 Revised:2017-09-27 Published:2017-12-15 Online:2017-12-15

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摘要： 目的：探讨微小RNA-10b（microRNA-10b）对宫颈癌细胞侵袭迁移能力的影响及机制。方法：实时定量聚合酶链反应（real-time PCR）检测microRNA-10b在不同宫颈癌细胞及正常宫颈上皮细胞中的表达情况；脂质体转染microRNA-10b模拟物（microRNA-10b mimics，即mimics组）及microRNA无关序列（microRNA-10b NC，即NC组）至Caski细胞，转染microRNA-10b抑制物（microRNA-10b inhibitor，即inhibitor组）及microRNA inhibitor无关序列（microRNA-10b NC inhibitor，即NC inhibitor组）至HeLa细胞，转染12 h后，real-time PCR法检测转染效率。采用Transwell试验和划痕试验分别检测microRNA-10b对Caski和HeLa细胞侵袭迁移能力的影响；real-time PCR及蛋白质印迹（Western blotting）检测各组细胞中胰岛素样生长因子1受体（IGF-1R）mRNA和蛋白表达水平；荧光素酶报告基因试验检测microRNA-10b对IGF-1R基因表达的调控机制。结果：MicroRNA-10b在宫颈癌细胞中表达水平低于正常宫颈上皮细胞（P＜0.05）。MicroRNA-10b表达增加能够降低Caski细胞的侵袭迁移能力（P＜0.05），抑制IGF-1R的mRNA和蛋白表达（均P＜0.05）；而microRNA-10b表达下降能够增强HeLa细胞的侵袭迁移能力（P＜0.05），增加IGF-1R的mRNA和蛋白表达（P均＜0.05）。在HEK-293T细胞中，microRNA-10b表达增加能够减弱含有IGF-1R基因3′非翻译区（3′-UTR）的报告质粒的荧光素酶活性（P＜0.05）。结论：在宫颈癌细胞中，microRNA-10b通过下调IGF-1R基因表达，抑制细胞侵袭迁移能力。
   

关键词: 微RNAs, 宫颈肿瘤, 肿瘤侵润, 细胞运动

Abstract: Objective：To investigate the role of microRNA-10b in the cervical carcinoma and explore its possible mechanism. Methods：The expression levels of microRNA-10b in cervical carcinoma cell lines and the normal cervical epithelial cells were detected by the real-time polymerase chain reaction (real-time PCR). The microRNA-10b mimics/NC was transfected into the Caski cells and the microRNA-10b inhibitor/NC inhibitor was transfected into the HeLa cells by the liposome. The Transwell assay and wound-healing assay were used to determine the effects of microRNA-10b on the invasion and migration ability in the Caski and HeLa cells. The mRNA and protein levels of insulin-like growth factor 1 receptor (IGF-1R) were analyzed by the real-time PCR and the Western blotting assay, respectively. Luciferase reporter assay was applied to detect the regulatory mechanism of microRNA-10b in the IGF-1R gene. Results：The expression levels of microRNA-10b in the cervical carcinoma cell lines were much lower than those in the normal cervical epithelial cells (P＜0.05). The up-regulation of microRNA-10b could decrease the invasion and migration ability of Caski cells (P＜0.05), and inhibit the mRNA and protein levels of IGF-1R (P＜0.05), while the downregulation of microRNA-10b could increase the invasion and migration ability of HeLa cells (P＜0.05), and increase the mRNA and protein levels of IGF-1R (P＜0.05). Besides, the up-regulation of microRNA-10b could decrease the luciferase activity of the reporter vector (containing the 5'untranslated region of IGF-1R gene) in the HEK-293T cells (P＜0.05). Conclusions：In cervical carcinoma cells, microRNA-10b could inhibit the cell invasion and migration capability, which may be the result of the down-regulation of IGF-1R gene.

Key words: MicroRNAs, Uterine cervical neoplasms, Neoplasm invasiveness, Cell movement