摘要
目的探究miRNA-204-5p通过靶向T细胞因子4(T cell factor 4,TCF-4)对宫颈癌Siha细胞增殖、侵袭和迁移的调节作用。方法miR-204mimic转染宫颈癌Siha细胞.实时荧光定量PCR检测miR-204及TCF-4的表达水平;生物信息学预测和荧光素酶报告实验检测miR-204和TCF-4的靶向关系:接着,miR-204mimic和pcDNA。TCF-4(pc-TCF-4)单独或一起转染Siha细胞,蛋白印记法检测TCF-4的表达;CCK8检测细胞活性;TransweH检测细胞侵袭能力;划痕实验检测细胞迁移能力。结果miR-204过表达能明显抑制宫颈癌Siha细胞TCF-4的表达(P<0.05);能显著降低野生型TCF-4荧光素酶活性(P<0.05);而TCF-4能明显减弱miR-204mimic对TCF-4表达的抑制作用(P<0.05);同时miR-204mimic可以显著抵消TCF-4过表达对宫颈癌细胞的增殖速率,侵袭细胞数,迁移的诱导作用(P<0.05)。结论miR-204能抑制宫颈癌细胞的增殖、侵袭和迁移,作用机制与靶向抑制TCF-4有关。
Objective To investigate the regulation of miRNA-204-5p on proliferation,invasion and migration of cervical cancer Siha cells by targeting TCF-4.Methods The expression levels of miR-204and TCF-4mRNA were detected by RT-PCR after Siha cells were transfected with miR- 204mimic.The targeting relationship between miR-204and TCF-4were determined by bioinformatics predicts and luciferase report assay.The protein level of TCF-4was detected by Western boltting after Siha cells were tranfected with miR-204and (or)pc-TCF-4.The cell viability was measured by CCK8assay.The metastatic ability was calculated by wound healing and Transwell assay.Results Overexpression of miR-204 could significantly reduce the level of TCF-4mRNA and wild-type TCF-4plasmid luciferase activity in cervical cancer Siha cells (P <0.05).The luciferase reporter assay indicated that there was miR-204-5p binding site on TCF-4.Meanwhile,mir-204mimic could significantly offset the inducing effect of TCF-4overexpresion on the proliferation,invasion,migration of cervical cancer cells (P <0.05).Conclusion miR-204inhibits the proliferation,invasion and migration of cervical cancer cells by targeting TCF4.
作者
曹锦慧
思美丽
CAO Jinhui;SI Meili(Department of Obstetrics and Gynecology,Yulin Xingyuan Hospital,Yulin,Shaanxi,719000,China;Department of Obstetrics and Gynecology,Yulin Second Hospital,Yulin,Shaanxi,719000,China)
出处
《医学分子生物学杂志》
CAS
2018年第6期373-378,共6页
Journal of Medical Molecular Biology
基金
陕西省科学技术研究发展计划项目(No.2012K16-09-03)。
