Department of Obstetrics & Gynaecology, The Chinese University of Hong Kong, Hong Kong, China.
Gynecol Oncol. 2013 Apr;129(1):199-208. doi: 10.1016/j.ygyno.2012.12.043. Epub 2013 Jan 9.
The purposes of this study were to identify aberrantly expressed miRNAs and investigate their pathogenic roles in cervical cancer.
miRNA expression was assessed in cervical cancer cell lines, micro-dissected normal cervical epithelium cells and primary cervical carcinoma by TaqMan RT-PCR. Spatial expression of miR-182 in cervical carcinoma and normal cervix was explored by in situ hybridization. HeLa xenograft mice model was used for evaluation of the effect on tumor growth of miR-182 inhibitor. Western blot, flow cytometry and gene expression analysis were used for identification of the functional role of miR-182 in HeLa cells.
Two up-regulated (miR-182 and -183) and nine down-regulated (miR-211, 145, 223, 150, 142-5p, 328, 195, 199b, 142-3p) microRNAs were consistently identified in cervical cancer cell lines. Further investigation confirmed the most up-regulated miRNA (miR-182) was significantly elevated in primary cervical carcinoma and discovered a significant correlation between the increased expression of miR-182 and advanced stages of cervical cancer. In HeLa xenograft mouse model, we demonstrated that inhibition of the miR-182 could exert the effect of tumor growth regression. Western blot, flow cytometry and pathway analysis for the HeLa cells with miR-182 over/down-expression in vitro showed that miR-182 was involved in apoptosis and cell cycle pathways, it also associated with the regulation of FOXO1.
Our findings indicated that miR-182 plays an onco-miRNA role in cervical cancer and its alteration is associated with cervical cancer pathogenesis by disrupting cell proliferation.
本研究旨在鉴定异常表达的 miRNAs，并研究其在宫颈癌中的致病作用。
采用 TaqMan RT-PCR 法检测宫颈癌细胞系、微切割正常宫颈上皮细胞和原发性宫颈癌中 miRNA 的表达。采用原位杂交法探讨 miR-182 在宫颈癌和正常宫颈中的空间表达。使用 HeLa 异种移植小鼠模型评估 miR-182 抑制剂对肿瘤生长的影响。采用 Western blot、流式细胞术和基因表达分析鉴定 miR-182 在 HeLa 细胞中的功能作用。
在宫颈癌细胞系中鉴定出两个上调（miR-182 和 miR-183）和九个下调（miR-211、145、223、150、142-5p、328、195、199b、142-3p）的 microRNAs。进一步的研究证实，在原发性宫颈癌中，最上调的 miRNA（miR-182）显著升高，并发现 miR-182 的表达增加与宫颈癌的晚期阶段之间存在显著相关性。在 HeLa 异种移植小鼠模型中，我们证明抑制 miR-182 可以发挥肿瘤生长消退的作用。体外对 miR-182 过表达/下调的 HeLa 细胞进行 Western blot、流式细胞术和通路分析表明，miR-182 参与凋亡和细胞周期通路，它还与 FOXO1 的调节有关。
我们的研究结果表明，miR-182 在宫颈癌中发挥癌基因 miRNA 的作用，其改变通过破坏细胞增殖与宫颈癌的发病机制有关。
