The effect of microRNA-129 on the migration and invasion in NSCLC cells and its mechanism

Purpose: To investigate the effect of microRNA-129 on the migration and invasion in (Non small cell lung cancer) NSCLC cells and involved mechanism. Materials and Methods: A549 cells were cultured and transfected with microRNA-129 (miR-129) mimic. Wound-healing assay and transwell assay were used to analyses the migration and invasion of A549 cells, respectively. The SOX4 expression changes were detected by qRT-PCR and Western blot. Dual-luciferase reporter assay was used to determine the binding site between miR-129 and the 3’-UTR of SOX4. A549 cells, miR-129 up-regulated A549 cells and miR-129 + SOX4 up-regulated A549 cells were treated with TGF-β in order to induce the epithelial-mesenchymal transition (EMT). The miR-129 and SOX4 after TGF-β treatment were measured. The EMT markers level changes were detected by western blot. Results: Up-regulation of miR-129 significantly inhibited the migration and invasion of A549 cells. miR-129 mimic transfection could reduce the mRNA and protein level of SOX4. Dual-luciferase reporter assay showed that miR-129 could bind to the 3’-UTR of SOX4. TGF-β induced EMT accompanied by the reduction of miR-129 and elevating of SOX4. Up-regulation of miR-129 could reverse the EMT marker changes caused by TGF-β, however, the SOX4 overexpression antagonized this effect of miR-129. Conclusions: miR-129 could suppress the migration and invasion of A549 cells. The potential mechanism may be that miR-129 inhibit EMT via targeting SOX4.研究目的：探讨微小RNA-129（microRNA-129，miR-129）对非小细胞肺癌（Non small cell lung cancer，NSCLC）细胞迁移与侵袭的影响及其潜在作用机制。 材料与方法：培养A549细胞并转染miR-129模拟物（mimic）。分别采用划痕愈合实验与Transwell实验分析A549细胞的迁移与侵袭能力。通过实时荧光定量PCR（qRT-PCR）与蛋白质印迹（Western blot）检测SOX4的表达变化。采用双荧光素酶报告基因实验验证miR-129与SOX4的3’-UTR的结合位点。将A549细胞、miR-129过表达A549细胞以及miR-129联合SOX4过表达A549细胞用转化生长因子-β（transforming growth factor-β，TGF-β）处理以诱导上皮间质转化（epithelial-mesenchymal transition，EMT），检测TGF-β处理后miR-129与SOX4的表达水平，并通过蛋白质印迹检测上皮间质转化标志物的表达变化。 实验结果：过表达miR-129可显著抑制A549细胞的迁移与侵袭能力。转染miR-129模拟物可降低SOX4的mRNA与蛋白表达水平。双荧光素酶报告基因实验证实，miR-129可结合SOX4的3’-UTR区域。TGF-β诱导的上皮间质转化伴随miR-129表达下调与SOX4表达上调。过表达miR-129可逆转TGF-β诱导的上皮间质转化标志物表达变化，而SOX4过表达则可拮抗miR-129的这一作用。 结论：miR-129可抑制A549细胞的迁移与侵袭能力，其潜在机制可能为通过靶向SOX4抑制上皮间质转化过程。