miR-145通过靶向AP4对非小细胞肺癌A549细胞迁移侵袭的影响

赵天增; 杨金华; 刘向前; 徐萌博

doi:10.3971/j.issn.1000-8578.2019.18.1160

miR-145通过靶向AP4对非小细胞肺癌A549细胞迁移侵袭的影响

miR-145 Attenuates Migration and Invasion of Non-small Cell Lung Cancer A549 Cells by Targeting AP4

摘要

摘要:
目的探讨miR-145对非小细胞肺癌迁移与侵袭能力的影响及其可能的作用机制。
方法 qRT-PCR法检测非小细胞肺癌组织中miR-145和激活增强子结合蛋白4（activating enhancer binding protein 4, AP4）mRNA的表达。A549细胞转染miR-145 mimic或者AP4 siRNA后，划痕实验与Transwell小室法分别检测A549细胞的迁移与侵袭能力，qRT-PCR法与免疫印迹法（Western blot）检测A549细胞中AP4 mRNA与蛋白表达水平；双荧光素酶报告基因法检测AP4 mRNA与miR-145的关系。
结果与癌旁正常组织相比，非小细胞肺癌组织中miR-145表达明显降低（P < 0.05），而AP4 mRNA和蛋白表达均明显升高（均P < 0.05）；与对照组相比，转染miR-145 mimic后，A549细胞中miR-145表达明显升高（P < 0.05），AP4 mRNA和蛋白水平明显降低（P < 0.05）；A549细胞的迁移数与侵袭能力均显著下降（均P < 0.05）；双荧光素酶报告基因法验证AP4 mRNA是miR-145的靶基因。A549细胞转染AP4 siRNA后，A549细胞的迁移数与侵袭数显著下降（均P < 0.05）。
结论上调miR-145能抑制A549细胞的迁移与侵袭能力，可能与其抑制靶基因AP4表达有关。

Abstract:
Objective To investigate the effect of miR-145 on the migration and invasion of non-small cell lung cancer (NSCLC) cells and possible mechanism.
Methods The expression of miR-145 and activating enhancer binding protein 4 (AP4) mRNA in NSCLC tissues were detected by qRT-PCR. After the transfection with miR-145 mimic or AP4 siRNA, the migration and invasion of A549 cells were examined by wound healing and Transwell assay, respectively; The mRNA and protein expressions of AP4 were detected by qRT-PCR and Western blot, respectively. The relationship between AP4 mRNA and miR-145 was detected by double luciferase reporter gene method.
Results Compared with adjacent normal tissues, the level of miR-145 was significantly decreased in NSCLC tissues (P < 0.05), however, the mRNA and protein levels of AP4 were significantly increased (P < 0.05). Compared with control group, the level of miR-145 was significantly increased (P < 0.05), the mRNA and protein levels of AP4 were significantly decreased (P < 0.05), and cells migration and invasion were significantly decreased (P < 0.05) in A549 cells after the transfection with miR-145 mimic. Moreover, AP4 was a direct target of miR-145. After the transfection with AP4 siRNA, the migration and invasion of A549 cells were significantly decreased (P < 0.05).
Conclusion miR-145 upregulation could efficiently attenuate the migration and invasion of A549 cells via inhibiting target gene AP4 expression.

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