Liu Ping, Wang Chao, Ma Chengbin, Wu Qiongwei, Zhang Wenying, Lao Guoying
Gynecology Department, Changning Maternity and Infant Health Hospital, No. 773, Wuyi Road, Changning District, Shanghai, 200051 China.
Cancer Cell Int. 2016 Sep 5;16(1):67. doi: 10.1186/s12935-016-0342-1. eCollection 2016.
To investigate the role of total cellular microRNA (miRNA) in regulating epithelial-to-mesenchymal transition (EMT) during human endometrial endometrioid adenocarcinoma (EEC).
A miRCURY LNA microRNA array was used to evaluate the miRNA profiles of human EEC tissues and corresponding nontumorous endometriums. An in vitro model of TGF-β induced EMT in HEC-1-A cells was used to investigate the role of miRNAs in the EEC during EMT. The expression of SMAD3, SMAD5, and a panel of EMT markers was detected by Western blot and quantitative PCR.
The results of miRNA profiling in human EEC tissues and corresponding nontumorous endometriums demonstrated that miR-23a expression was down-regulated. Using bioinformatics, we identified SMAD3 or SMAD5 maybe as a predicted target of miR-23a. The results of luciferase reporter assay showed miR-23a directly targets and down-regulates human SMAD3 protein levels, not SMAD5 protein levels. Furthermore, overexpression of miR-23a in HEC-1-A cells increased E-cadherin expression and decreased the expression of vimentin and alpha smooth muscle actin, markers of mesenchymal cellular phenotype.
Our data provide firm evidence of a role for miR-23a in the direct regulation of EMT through its targeting of SMAD3. Due to its ability to repress the EMT, miR-23a may be a novel target for EER therapeutic intervention.
研究全细胞微小RNA（miRNA）在人类子宫内膜样腺癌（EEC）上皮-间质转化（EMT）过程中的调控作用。
使用miRCURY LNA微小RNA芯片评估人类EEC组织及相应的非肿瘤性子宫内膜的miRNA谱。利用TGF-β诱导HEC-1-A细胞发生EMT的体外模型，研究miRNA在EEC的EMT过程中的作用。通过蛋白质免疫印迹法和定量PCR检测SMAD3、SMAD5以及一组EMT标志物的表达。
人类EEC组织及相应的非肿瘤性子宫内膜的miRNA谱分析结果表明，miR-23a表达下调。通过生物信息学分析，我们确定SMAD3或SMAD5可能是miR-23a的预测靶标。荧光素酶报告基因检测结果显示，miR-23a直接靶向并下调人类SMAD3蛋白水平，而非SMAD5蛋白水平。此外，在HEC-1-A细胞中过表达miR-23a可增加E-钙黏蛋白的表达，并降低波形蛋白和α平滑肌肌动蛋白的表达，这两种蛋白是间充质细胞表型的标志物。
我们的数据提供了确凿证据，证明miR-23a通过靶向SMAD3直接调控EMT。由于其抑制EMT的能力，miR-23a可能是EEC治疗干预的新靶点。
