摘要
目的探讨microRNA-182(miR-182)在宫颈癌细胞增殖中的作用及其机制。方法使用脂质体介导的瞬时转染法增加或降低宫颈癌细胞系He La和Si Ha细胞中miR-182水平,利用CCK-8法和平板克隆形成实验观察miR-182对宫颈癌细胞增殖能力的影响,使用生物信息学预测、实时定量PCR和双荧光素酶报告基因等实验明确miR-182对腺瘤性息肉病基因(APC)的转录后基因表达调控的作用以及对Wnt下游经典信号分子c-Myc和细胞周期蛋白D1(cyclin D1)水平的影响。结果过表达miR-182明显促进宫颈癌细胞的增殖,抑制miR-182的表达则显著抑制肿瘤细胞的增殖;过表达miR-182抑制宫颈癌细胞中APC的表达,调控miR-182的表达水平影响肿瘤细胞中经典Wnt信号通路下游的表达。结论 miR-182通过抑制APC的表达激活经典Wnt信号通路的活性,提高宫颈癌细胞增殖能力。
Objective To investigate the role and mechanism of microRNA-182 (miR-182) in the proliferation of cervical cancer cells. Methods With liposome-mediated transient transfection method, the level of miR-182 in HeLa and SiHa cells was increased or decreased. CCK-8 assay and colony formation assay were used to observe the effect of miR-182 on the proliferation of cervical cancer cells. Using bioinformatics predictions, real-time quantitative PCR, and dual luciferase reporter assay, we clarified the role of miR-182 in posttranscriptional regulation of adenomatous polyposis coli (APC) gene and its effect on the downstream molecules (c-Myc and cyclin D1) of Writ singling pathway. Results Up-regulation of miR-182 significantly promoted the proliferation of cervical cancer cells, while down-regulation of miR-182 significantly inhibited the proliferation of cervical cancer cells. Over-expression of miR-182 inhibited the expression of APC gene in cervical cancer cells and the regulation of miR-182 affected the expression of canonical Writ signaling pathway downstream molecules in cervical cancer cells. Conclusion The miR-182 stimulates canonical Wnt signaling pathway by targeting APC gene and enhances the proliferation of cervical cancer cells.
作者
李佩
胡静
张莹
李剑平
党运芝
张瑞
魏丽春
石梅
LIPei;HU Jing;ZHANGYing;LI Jianping;DANG Yunzhi;ZHANG Rui;WEI Lichun;SHI Mei(Department of Radiation Oncology, Xijing Hospital;Department of Biochemistry and Molecular Biology, Air Force Military Medical University, Xi'an 710032, China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2018年第2期148-153,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金(81272346)