Cheng Yuan, Guo Yanli, Zhang Youyi, You Ke, Li Zijian, Geng Li
Department of Gynecology and Obstetrics, Peking University Third Hospital, Beijing, 100191, China.
Institute of Vascular Medicine, Peking University Third Hospital, Beijing Key Laboratory of Cardiovasicular Receptors Research, Key Laboratory of Cardiovascular Molecular Biology and Regulatory Peptides, Ministry of Health and Key Laboratory of Molecular Cardiovascular Sciences, Ministry of Education, Beijing, 100191, China.
J Exp Clin Cancer Res. 2016 Jan 15;35:11. doi: 10.1186/s13046-016-0290-6.
MicroRNA-106b (miR-106b) was recently identified as an oncogene participating in cancer progression. Transforming growth factor β1(TGF-β1) is an indispensable cytokine regulating the local microenvironment, thereby promoting cervical cancer progression. However, the roles of miR-106b in cervical carcinoma progression and TGF-β1-involvement in the tumorigenesis of cervical cancer remain unknown.
The expression of miR-106b in human cervical specimens was detected by real-time PCR analysis and in situ hybridization assay. The effect of miR-106b on cell migration was analyzed by scratch and transwell assays. TGF-β1 was used to induce cell migration. The expression of the miR-106b target gene DAB2 in human cervical tissues and cell lines were measured by qRT-PCR, western blot and immunohistochemistry. Dual-luciferase reporter assay was used to identify DAB2 as a miR-106b-directed target gene.
miR-106b was frequently up-regulated in human cervical carcinoma specimens and cervical cancer cell lines. Over-expression of miR-106b significantly promoted HeLa and SiHa cells migration. Likewise, inhibition of miR-106b decreased HeLa and SiHa cells migration. The multifunctional cytokine TGF-β facilitates metastasis in cervical carcinoma. miR-106b inhibitor treatment decreased the TGF-β1-stimulated migration of HeLa and SiHa cells. DAB2, a predicted target gene of miR-106b, was inhibited by TGF-β1 partly through miR-106b and was involved in TGF-β1-induced cervical cancer cell migration. The expression of DAB2 was low in cervical cancer tissues, and negatively correlated with miR-106b expression. Finally, DAB2 was identified as a miR-106b-directed target gene by dual-luciferase reporter assay.
Our data suggest that the TGF-β1/miR-106b/DAB2 axis may be involved in the pathogenesis of cervical carcinoma.
微小RNA-106b（miR-106b）最近被鉴定为参与癌症进展的致癌基因。转化生长因子β1（TGF-β1）是调节局部微环境从而促进宫颈癌进展的不可或缺的细胞因子。然而，miR-106b在宫颈癌进展中的作用以及TGF-β1参与宫颈癌发生的机制仍不清楚。
通过实时PCR分析和原位杂交检测人宫颈标本中miR-106b的表达。通过划痕试验和Transwell试验分析miR-106b对细胞迁移的影响。用TGF-β1诱导细胞迁移。通过qRT-PCR、蛋白质免疫印迹和免疫组织化学检测人宫颈组织和细胞系中miR-106b靶基因DAB2的表达。采用双荧光素酶报告基因检测法鉴定DAB2为miR-106b直接作用的靶基因。
miR-106b在人宫颈癌标本和宫颈癌细胞系中经常上调。miR-106b的过表达显著促进HeLa和SiHa细胞迁移。同样，抑制miR-106b可降低HeLa和SiHa细胞迁移。多功能细胞因子TGF-β促进宫颈癌转移。miR-106b抑制剂处理可降低TGF-β1刺激的HeLa和SiHa细胞迁移。DAB2是miR-106b的预测靶基因，部分通过miR-106b被TGF-β1抑制，并参与TGF-β1诱导的宫颈癌细胞迁移。DAB2在宫颈癌组织中的表达较低，且与miR-106b表达呈负相关。最后，通过双荧光素酶报告基因检测法鉴定DAB2为miR-106b直接作用的靶基因。
我们的数据表明，TGF-β1/miR-106b/DAB2轴可能参与宫颈癌的发病机制。
